Macrocyclic inhibitors of hepatitis C virus

ABSTRACT

Inhibitors of HCV replication of formula (I) 
     
       
         
         
             
             
         
       
         
         and the N-oxides, salts, and stereoisomers, wherein 
         each dashed line represents an optional double bond; 
         X is N, CH and where X bears a double bond it is C; 
         R 1  is —OR 7 , —NH—SO 2 R 8 ; 
         R 2  is hydrogen, and where X is C or CH, R 2  may also be C 1-6 alkyl; 
         R 3  is hydrogen, C 1-6 alkyl, C 1-6 alkoxyC 1-6 alkyl, C 3-7 cycloalkyl; 
         R 4  is aryl or Het; n is 3, 4, 5, or 6; 
         R 5  is halo, C 1-6 alkyl, hydroxy, C 1-6 alkoxy, phenyl, or Het; 
         R 6  is C 1-6 alkoxy, or dimethylamino; 
         R 7  is hydrogen; aryl; Het; C 3-7 cycloalkyl optionally substituted with C 1-6 alkyl; or C 1-6 alkyl optionally substituted with C 3-7 cycloalkyl, aryl or with Het; 
         R 8  is aryl; Het; C 3-7 cycloalkyl optionally substituted with C 1-6 alkyl; or C 1-6 alkyl optionally substituted with C 3-7 cycloalkyl, aryl or with Het; 
         aryl is phenyl optionally substituted with one, two or three substituents; 
         Het is a 5 or 6 membered saturated, partially unsaturated or completely unsaturated heterocyclic ring containing 1 to 4 heteroatoms selected from nitrogen, oxygen and sulfur, and being optionally substituted with one, two or three substituents; 
         pharmaceutical compositions containing compounds (I) and processes for preparing compounds (I). Bioavailable combinations of the inhibitors of HCV of formula (I) with ritonavir are also provided.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/412,997, filed Mar. 6, 2012, which is a divisional of U.S.application Ser. No. 13/197,226, filed Aug. 3, 2011, now U.S. Pat. No.8,153,800, which is a divisional of U.S. application Ser. No.11/632,102, now U.S. Pat. No. 8,148,399, filed Jan. 10, 2007, which isthe National Stage Entry of International Application No.PCT/EP2006/064820, filed Jul. 28, 2006, which claims the benefit ofEuropean Application Nos. 06101280.3, filed Feb. 3, 2006, 05107417.7,filed Aug. 11, 2005, and 05107074.6, filed Jul. 29, 2005, the entiretiesof which are incorporated herein by reference.

The present invention is concerned with macrocylic compounds havinginhibitory activity on the replication of the hepatitis C virus (HCV).It further concerns compositions comprising these compounds as activeingredients as well as processes for preparing these compounds andcompositions.

Hepatitis C virus is the leading cause of chronic liver diseaseworldwide and has become a focus of considerable medical research. HCVis a member of the Flaviviridae family of viruses in the hepacivirusgenus, and is closely related to the flavivirus genus, which includes anumber of viruses implicated in human disease, such as dengue virus andyellow fever virus, and to the animal pestivirus family, which includesbovine viral diarrhea virus (BVDV). HCV is a positive-sense,single-stranded RNA virus, with a genome of around 9,600 bases. Thegenome comprises both 5′ and 3′ untranslated regions which adopt RNAsecondary structures, and a central open reading frame that encodes asingle polyprotein of around 3,010-3,030 amino acids. The polyproteinencodes ten gene products which are generated from the precursorpolyprotein by an orchestrated series of co- and posttranslationalendoproteolytic cleavages mediated by both host and viral proteases. Theviral structural proteins include the core nucleocapsid protein, and twoenvelope glycoproteins E1 and E2. The non-structural (NS) proteinsencode some essential viral enzymatic functions (helicase, polymerase,protease), as well as proteins of unknown function. Replication of theviral genome is mediated by an RNA-dependent RNA polymerase, encoded bynon-structural protein 5b (NS5B). In addition to the polymerase, theviral helicase and protease functions, both encoded in the bifunctionalNS3 protein, have been shown to be essential for replication of HCV RNA.In addition to the NS3 serine protease, HCV also encodes ametalloproteinase in the NS2 region.

Following the initial acute infection, a majority of infectedindividuals develop chronic hepatitis because HCV replicatespreferentially in hepatocytes but is not directly cytopathic. Inparticular, the lack of a vigorous T-lymphocyte response and the highpropensity of the virus to mutate appear to promote a high rate ofchronic infection. Chronic hepatitis can progress to liver fibrosisleading to cirrhosis, end-stage liver disease, and HCC (hepatocellularcarcinoma), making it the leading cause of liver transplantations.

There are 6 major HCV genotypes and more than 50 subtypes, which aredifferently distributed geographically. HCV type 1 is the predominantgenotype in Europe and the US. The extensive genetic heterogeneity ofHCV has important diagnostic and clinical implications, perhapsexplaining difficulties in vaccine development and the lack of responseto therapy.

Transmission of HCV can occur through contact with contaminated blood orblood products, for example following blood transfusion or intravenousdrug use. The introduction of diagnostic tests used in blood screeninghas led to a downward trend in post-transfusion HCV incidence. However,given the slow progression to the end-stage liver disease, the existinginfections will continue to present a serious medical and economicburden for decades.

Current HCV therapies are based on (pegylated) interferon-alpha (IFN-α)in combination with ribavirin. This combination therapy yields asustained virologic response in more than 40% of patients infected bygenotype 1 viruses and about 80% of those infected by genotypes 2 and 3.Beside the limited efficacy on HCV type 1, this combination therapy hassignificant side effects and is poorly tolerated in many patients. Majorside effects include influenza-like symptoms, hematologic abnormalities,and neuropsychiatric symptoms. Hence there is a need for more effective,convenient and better tolerated treatments.

Recently, two peptidomimetic HCV protease inhibitors have gainedattention as clinical candidates, namely BILN-2061 disclosed inWO00/59929 and VX-950 disclosed in WO03/87092. A number of similar HCVprotease inhibitors have also been disclosed in the academic and patentliterature. It has already become apparent that the sustainedadministration of BILN-2061 or VX-950 selects HCV mutants which areresistant to the respective drug, so called drug escape mutants. Thesedrug escape mutants have characteristic mutations in the HCV proteasegenome, notably D168V, D168A and/or A156S. Accordingly, additional drugswith different resistance patterns are required to provide failingpatients with treatment options, and combination therapy with multipledrugs is likely to be the norm in the future, even for first linetreatment.

Experience with HIV drugs, and HIV protease inhibitors in particular,has further emphasized that sub-optimal pharmacokinetics and complexdosage regimes quickly result in inadvertent compliance failures. Thisin turn means that the 24 hour trough concentration (minimum plasmaconcentration) for the respective drugs in an HIV regime frequentlyfalls below the IC₉₀ or ED₉₀ threshold for large parts of the day. It isconsidered that a 24 hour trough level of at least the IC₅₀, and morerealistically, the IC₉₀ or ED₉₀, is essential to slow down thedevelopment of drug escape mutants. Achieving the necessarypharmacokinetics and drug metabolism to allow such trough levelsprovides a stringent challenge to drug design. The strong peptidomimeticnature of prior art HCV protease inhibitors, with multiple peptide bondsposes pharmacokinetic hurdles to effective dosage regimes.

There is a need for HCV inhibitors which may overcome the disadvantagesof current HCV therapy such as side effects, limited efficacy, theemerging of resistance, and compliance failures.

The present invention concerns HCV inhibitors which are superior in oneor more of the following pharmacological related properties, i.e.potency, decreased cytotoxicity, improved pharmacokinetics, improvedresistance profile, acceptable dosage and pill burden.

In addition, the compounds of the present invention have relatively lowmolecular weight and are easy to synthesize, starting from startingmaterials that are commercially available or readily available throughart-known synthesis procedures.

WO05/010029 discloses aza-peptide macrocyclic Hepatitis C serineprotease inhibitors, pharmaceutical compositions comprising theaforementioned compounds for administration to a subject suffering fromHCV infection, and methods of treating an HCV infection in a subject byadministering a pharmaceutical composition comprising the saidcompounds.

The present invention concerns inhibitors of HCV replication, which canbe represented by formula (I):

-   and the N-oxides, salts, and stereoisomers thereof, wherein-   each dashed line (represented by - - - - -) represents an optional    double bond;-   X is N, CH and where X bears a double bond it is C;-   R¹ is —OR⁷, —NH—SO₂R⁸;-   R² is hydrogen, and where X is C or CH, R² may also be C₁₋₆alkyl;-   R³ is hydrogen, C₁₋₆alkyl, C₁₋₆alkoxyC₁₋₆alkyl, C₃₋₇cycloalkyl;-   R⁴ is aryl or Het;-   n is 3, 4, 5, or 6;-   R⁵ represents halo, C₁₋₆alkyl, hydroxy, C₁₋₆alkoxy,    polyhaloC₁₋₆alkyl, phenyl, or Het;-   R⁶ represents C₁₋₆alkoxy, mono- or diC₁₋₆alkylamino;-   R⁷ is hydrogen; aryl; Het; C₃₋₇cycloalkyl optionally substituted    with C₁₋₆alkyl; or C₁₋₆alkyl optionally substituted with    C₃₋₇cycloalkyl, aryl or with Het;-   R⁸ is aryl; Het; C₃₋₇cycloalkyl optionally substituted with    C₁₋₆alkyl; or C₁₋₆alkyl optionally substituted with C₃₋₇cycloalkyl,    aryl or with Het;-   aryl as a group or part of a group is phenyl optionally substituted    with one, two or three substituents selected from halo, hydroxy,    nitro, cyano, carboxyl, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆alkoxyC₁₋₆alkyl,    C₁₋₆alkylcarbonyl, amino, mono- or di-C₁₋₆alkyl-amino, azido,    mercapto, polyhaloC₁₋₆alkyl, polyhaloC₁₋₆alkoxy, C₃₋₇cycloalkyl,    pyrrolidinyl, piperidinyl, piperazinyl, 4-C₁₋₆alkyl-piperazinyl,    4-C₁₋₆alkylcarbonyl-piperazinyl, and morpholinyl; wherein the    morpholinyl and piperidinyl groups may be optionally substituted    with one or with two C₁₋₆alkyl radicals;-   Het as a group or part of a group is a 5 or 6 membered saturated,    partially unsaturated or completely unsaturated heterocyclic ring    containing 1 to 4 heteroatoms each independently selected from    nitrogen, oxygen and sulfur, said heterocyclic ring being optionally    condended with a benzene ring; and Het as a whole being optionally    substituted with one, two or three substituents each independently    selected from the group consisting of halo, hydroxy, nitro, cyano,    carboxyl, C₁₋₆alkyl, C₁₋₆alkoxy, C₁₋₆alkoxyC₁₋₆alkyl,    C₁₋₆alkylcarbonyl, amino, mono- or di-C₁₋₆alkylamino, azido,    mercapto, polyhaloC₁₋₆alkyl, polyhaloC₁₋₆alkoxy, C₃₋₇cycloalkyl,    pyrrolidinyl, piperidinyl, piperazinyl, 4-C₁₋₆alkylpiperazinyl,    4-C₁₋₆alkylcarbonylpiperazinyl, and morpholinyl; wherein the    morpholinyl and piperidinyl groups may be optionally substituted    with one or with two C₁₋₆alkyl radicals.

The invention further relates to methods for the preparation of thecompounds of formula (I), the N-oxides, addition salts, quaternaryamines, metal complexes, and stereochemically isomeric forms thereof,their intermediates, and the use of the intermediates in the preparationof the compounds of formula (I).

The invention relates to the compounds of formula (I) per se, theN-oxides, addition salts, quaternary amines, metal complexes, andstereochemically isomeric forms thereof, for use as a medicament. Theinvention further relates to pharmaceutical compositions comprising acarrier and an anti-virally effective amount of a compound of formula(I) as specified herein. The pharmaceutical compositions may comprisecombinations of the aforementioned compounds with other anti-HCV agents.The invention further relates to the aforementioned pharmaceuticalcompositions for administration to a subject suffering from HCVinfection.

The invention also relates to the use of a compound of formula (I), or aN-oxide, addition salt, quaternary amine, metal complex, orstereochemically isomeric forms thereof, for the manufacture of amedicament for inhibiting HCV replication. Or the invention relates to amethod of inhibiting HCV replication in a warm-blooded animal saidmethod comprising the administration of an effective amount of acompound of formula (I), or a prodrug, N-oxide, addition salt,quaternary amine, metal complex, or stereochemically isomeric formsthereof.

As used in the foregoing and hereinafter, the following definitionsapply unless otherwise noted.

The term halo is generic to fluoro, chloro, bromo and iodo.

The term “polyhaloC₁₋₆alkyl” as a group or part of a group, e.g. inpolyhaloC₁₋₆alkoxy, is defined as mono- or polyhalo substitutedC₁₋₆alkyl, in particular C₁₋₆alkyl substituted with up to one, two,three, four, five, six, or more halo atoms, such as methyl or ethyl withone or more fluoro atoms, for example, difluoromethyl, trifluoromethyl,trifluoro-ethyl. Preferred is trifluoromethyl. Also included areperfluoroC₁₋₆alkyl groups, which are C₁₋₆alkyl groups wherein allhydrogen atoms are replaced by fluoro atoms, e.g. pentafluoroethyl. Incase more than one halogen atom is attached to an alkyl group within thedefinition of polyhaloC₁₋₆alkyl, the halogen atoms may be the same ordifferent.

As used herein “C₁₋₄alkyl” as a group or part of a group definesstraight or branched chain saturated hydrocarbon radicals having from 1to 4 carbon atoms such as for example methyl, ethyl, 1-propyl, 2-propyl,1-butyl, 2-butyl, 2-methyl-1-propyl; “C₁₋₆alkyl” encompasses C₁₋₄alkylradicals and the higher homologues thereof having 5 or 6 carbon atomssuch as, for example, 1-pentyl, 2-pentyl, 3-pentyl, 1-hexyl, 2-hexyl,2-methyl-1-butyl, 2-methyl-1-pentyl, 2-ethyl-1-butyl, 3-methyl-2-pentyl,and the like. Of interest amongst C₁₋₆alkyl is C₁₋₄alkyl.

The term “C₂₋₆alkenyl” as a group or part of a group defines straightand branched chained hydrocarbon radicals having saturated carbon-carbonbonds and at least one double bond, and having from 2 to 6 carbon atoms,such as, for example, ethenyl (or vinyl), 1-propenyl, 2-propenyl (orallyl), 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-2-propenyl,2-pentenyl, 3-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl,2-methyl-2-butenyl, 2-methyl-2-pentenyl and the like. Of interestamongst C₂₋₆alkenyl is C₂₋₄alkenyl.

The term “C₂₋₆alkynyl” as a group or part of a group defines straightand branched chained hydrocarbon radicals having saturated carbon-carbonbonds and at least one triple bond, and having from 2 to 6 carbon atoms,such as, for example, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl,2-butynyl, 3-butynyl, 2-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl andthe like. Of interest amongst C₂₋₆alkynyl is C₂₋₄alkynyl.

C₃₋₇cycloalkyl is generic to cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and cycloheptyl.

C₁₋₆alkanediyl defines bivalent straight and branched chain saturatedhydrocarbon radicals having from 1 to 6 carbon atoms such as, forexample, methylene, ethylene, 1,3-propanediyl, 1,4-butanediyl,1,2-propanediyl, 2,3-butanediyl, 1,5-pentanediyl, 1,6-hexanediyl and thelike. Of interest amongst C₁₋₆alkanediyl is C₁₋₄alkanediyl.

C₁₋₆alkoxy means C₁₋₆alkyloxy wherein C₁₋₆alkyl is as defined above.

As used herein before, the term (═O) or oxo forms a carbonyl moiety whenattached to a carbon atom, a sulfoxide moiety when attached to a sulfuratom and a sulfonyl moiety when two of said terms are attached to asulfur atom. Whenever a ring or ring system is substituted with an oxogroup, the carbon atom to which the oxo is linked is a staturatedcarbon.

The radical Het is a heterocycle as specified in this specification andclaims. Preferred amongst the Het radicals are those that aremonocyclic.

Examples of Het comprise, for example, pyrrolidinyl, piperidinyl,morpholinyl, thiomorpholinyl, piperazinyl, pyrrolyl, pyrazolyl,imidazolyl, oxazolyl, isoxazolyl, thiazinolyl, isothiazinolyl,thiazolyl, isothiazolyl, oxadiazolyl, thiadiazolyl, triazolyl (including1,2,3-triazolyl, 1,2,4-triazolyl), tetrazolyl, furanyl, thienyl,pyridyl, pyrimidyl, pyridazinyl, triazinyl, and the like. Of interestamongst the Het radicals are those which are non-saturated, inparticular those having an aromatic character. Of further interest arethose Het radicals having one or two nitrogens.

Each of the Het radicals mentioned in this and the following paragraphmay be optionally substituted with the number and kind of substituentsmentioned in the definitions of the compounds of formula (I) or any ofthe subgroups of compounds of formula (I). Some of the Het radicalsmentioned in this and the following paragraph may be substituted withone, two or three hydroxy substituents. Such hydroxy substituted ringsmay occur as their tautomeric forms bearing keto groups. For example a3-hydroxypyridazine moiety can occur in its tautomeric form2H-pyridazin-3-one. Where Het is piperazinyl, it preferably issubstituted in its 4-position by a substituent linked to the 4-nitrogenwith a carbon atom, e.g. 4-C₁₋₆alkyl, 4-polyhaloC₁₋₆alkyl,C₁₋₆alkoxyC₁₋₆alkyl, C₁₋₆alkylcarbonyl, C₃₋₇cycloalkyl.

Interesting Het radicals comprise, for example pyrrolidinyl,piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, pyrrolyl,pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl,oxadiazolyl, thiadiazolyl, triazolyl (including 1,2,3-triazolyl,1,2,4-triazolyl), tetrazolyl, furanyl, thienyl, pyridyl, pyrimidyl,pyridazinyl, pyrazolyl, triazinyl, or any of such heterocycles condensedwith a benzene ring, such as indolyl, indazolyl (in particular1H-indazolyl), indolinyl, quinolinyl, tetrahydroquinolinyl (inparticular 1,2,3,4-tetrahydroquinolinyl), isoquinolinyl,tetrahydroisoquinolinyl (in particular 1,2,3,4-tetrahydroisoquinolinyl),quinazolinyl, phthalazinyl, benzimidazolyl, benzoxazolyl,benzisoxazolyl, benzothiazolyl, benzoxadiazolyl, benzothiadiazolyl,benzofuranyl, benzothienyl.

The Het radicals pyrrolidinyl, piperidinyl, morpholinyl,thiomorpholinyl, piperazinyl, 4-substituted piperazinyl preferably arelinked via their nitrogen atom (i.e. 1-pyrrolidinyl, 1-piperidinyl,4-thiomorpholinyl, 4-morpholinyl, 1-piperazinyl, 4-substituted1-piperazinyl).

It should be noted that the radical positions on any molecular moietyused in the definitions may be anywhere on such moiety as long as it ischemically stable.

Radicals used in the definitions of the variables include all possibleisomers unless otherwise indicated. For instance pyridyl includes2-pyridyl, 3-pyridyl and 4-pyridyl; pentyl includes 1-pentyl, 2-pentyland 3-pentyl.

When any variable occurs more than one time in any constituent, eachdefinition is independent.

Whenever used hereinafter, the term “compounds of formula (I)”, or “thepresent compounds” or similar terms, it is meant to include thecompounds of formula (I), their prodrugs, N-oxides, addition salts,quaternary amines, metal complexes, and stereochemically isomeric forms.One embodiment comprises the compounds of formula (I) or any subgroup ofcompounds of formula (I) specified herein, as well as the N-oxides,salts, as the possible stereoisomeric forms thereof. Another embodimentcomprises the compounds of formula (I) or any subgroup of compounds offormula (I) specified herein, as well as the salts as the possiblestereoisomeric forms thereof.

The compounds of formula (I) have several centers of chirality and existas stereochemically isomeric forms. The term “stereochemically isomericforms” as used herein defines all the possible compounds made up of thesame atoms bonded by the same sequence of bonds but having differentthree-dimensional structures which are not interchangeable, which thecompounds of formula (I) may possess.

With reference to the instances where (R) or (S) is used to designatethe absolute configuration of a chiral atom within a substituent, thedesignation is done taking into consideration the whole compound and notthe substituent in isolation.

Unless otherwise mentioned or indicated, the chemical designation of acompound encompasses the mixture of all possible stereochemicallyisomeric forms, which said compound may possess. Said mixture maycontain all diastereomers and/or enantiomers of the basic molecularstructure of said compound. All stereochemically isomeric forms of thecompounds of the present invention both in pure form or mixed with eachother are intended to be embraced within the scope of the presentinvention.

Pure stereoisomeric forms of the compounds and intermediates asmentioned herein are defined as isomers substantially free of otherenantiomeric or diastereomeric forms of the same basic molecularstructure of said compounds or intermediates. In particular, the term“stereoisomerically pure” concerns compounds or intermediates having astereoisomeric excess of at least 80% (i.e. minimum 90% of one isomerand maximum 10% of the other possible isomers) up to a stereoisomericexcess of 100% (i.e. 100% of one isomer and none of the other), more inparticular, compounds or intermediates having a stereoisomeric excess of90% up to 100%, even more in particular having a stereoisomeric excessof 94% up to 100% and most in particular having a stereoisomeric excessof 97% up to 100%. The terms “enantiomerically pure” and“diastereomerically pure” should be understood in a similar way, butthen having regard to the enantiomeric excess, and the diastereomericexcess, respectively, of the mixture in question.

Pure stereoisomeric forms of the compounds and intermediates of thisinvention may be obtained by the application of art-known procedures.For instance, enantiomers may be separated from each other by theselective crystallization of their diastereomeric salts with opticallyactive acids or bases. Examples thereof are tartaric acid,dibenzoyltartaric acid, ditoluoyltartaric acid and camphosulfonic acid.Alternatively, enantiomers may be separated by chromatographictechniques using chiral stationary phases. Said pure stereochemicallyisomeric forms may also be derived from the corresponding purestereochemically isomeric forms of the appropriate starting materials,provided that the reaction occurs stereospecifically. Preferably, if aspecific stereoisomer is desired, said compound will be synthesized bystereospecific methods of preparation. These methods will advantageouslyemploy enantiomerically pure starting materials.

The diastereomeric racemates of the compounds of formula (I) can beobtained separately by conventional methods. Appropriate physicalseparation methods that may advantageously be employed are, for example,selective crystallization and chromatography, e.g. columnchromatography.

For some of the compounds of formula (I), their prodrugs, N-oxides,salts, solvates, quaternary amines, or metal complexes, and theintermediates used in the preparation thereof, the absolutestereochemical configuration was not experimentally determined A personskilled in the art is able to determine the absolute configuration ofsuch compounds using art-known methods such as, for example, X-raydiffraction.

The present invention is also intended to include all isotopes of atomsoccurring on the present compounds. Isotopes include those atoms havingthe same atomic number but different mass numbers. By way of generalexample and without limitation, isotopes of hydrogen include tritium anddeuterium. Isotopes of carbon include C-13 and C-14.

The term “prodrug” as used throughout this text means thepharmacologically acceptable derivatives such as esters, amides andphosphates, such that the resulting in vivo biotransformation product ofthe derivative is the active drug as defined in the compounds of formula(I). The reference by Goodman and Gilman (The Pharmacological Basis ofTherapeutics, 8^(th) ed, McGraw-Hill, Int. Ed. 1992, “Biotransformationof Drugs”, p 13-15) describing prodrugs generally is herebyincorporated. Prodrugs preferably have excellent aqueous solubility,increased bioavailability and are readily metabolized into the activeinhibitors in vivo. Prodrugs of a compound of the present invention maybe prepared by modifying functional groups present in the compound insuch a way that the modifications are cleaved, either by routinemanipulation or in vivo, to the parent compound.

Preferred are pharmaceutically acceptable ester prodrugs that arehydrolysable in vivo and are derived from those compounds of formula (I)having a hydroxy or a carboxyl group. An in vivo hydrolysable ester isan ester, which is hydrolysed in the human or animal body to produce theparent acid or alcohol. Suitable pharmaceutically acceptable esters forcarboxy include C₁₋₆alkoxymethyl esters for example methoxy-methyl,C₁₋₆alkanoyloxymethyl esters for example pivaloyloxymethyl, phthalidylesters, C₃₋₈cycloalkoxycarbonyloxyC₁₋₆alkyl esters for example1-cyclohexylcarbonyl-oxyethyl; 1,3-dioxolen-2-onylmethyl esters forexample 5-methyl-1,3-dioxolen-2-onylmethyl; andC₁₋₆alkoxycarbonyloxyethyl esters for example 1-methoxycarbonyl-oxyethylwhich may be formed at any carboxy group in the compounds of thisinvention.

An in vivo hydrolysable ester of a compound of the formula (I)containing a hydroxy group includes inorganic esters such as phosphateesters and α-acyloxyalkyl ethers and related compounds which as a resultof the in vivo hydrolysis of the ester breakdown to give the parenthydroxy group. Examples of α-acyloxyalkyl ethers include acetoxy-methoxyand 2,2-dimethylpropionyloxy-methoxy. A selection of in vivohydrolysable ester forming groups for hydroxy include alkanoyl, benzoyl,phenylacetyl and substituted benzoyl and phenylacetyl, alkoxycarbonyl(to give alkyl carbonate esters), dialkylcarbamoyl andN-(dialkylaminoethyl)-N-alkylcarbamoyl (to give carbamates),dialkylaminoacetyl and carboxyacetyl. Examples of substituents onbenzoyl include morpholino and piperazino linked from a ring nitrogenatom via a methylene group to the 3- or 4-position of the benzoyl ring.

For therapeutic use, salts of the compounds of formula (I) are thosewherein the counter-ion is pharmaceutically acceptable. However, saltsof acids and bases which are non-pharmaceutically acceptable may alsofind use, for example, in the preparation or purification of apharmaceutically acceptable compound. All salts, whetherpharmaceutically acceptable or not are included within the ambit of thepresent invention.

The pharmaceutically acceptable acid and base addition salts asmentioned hereinabove are meant to comprise the therapeutically activenon-toxic acid and base addition salt forms which the compounds offormula (I) are able to form. The pharmaceutically acceptable acidaddition salts can conveniently be obtained by treating the base formwith such appropriate acid. Appropriate acids comprise, for example,inorganic acids such as hydrohalic acids, e.g. hydrochloric orhydrobromic acid, sulfuric, nitric, phosphoric and the like acids; ororganic acids such as, for example, acetic, propanoic, hydroxyacetic,lactic, pyruvic, oxalic (i.e. ethanedioic), malonic, succinic (i.e.butanedioic acid), maleic, fumaric, malic (i.e. hydroxybutanedioicacid), tartaric, citric, methanesulfonic, ethanesulfonic,benzenesulfonic, p-toluenesulfonic, cyclamic, salicylic,p-aminosalicylic, pamoic and the like acids.

Conversely said salt forms can be converted by treatment with anappropriate base into the free base form.

The compounds of formula (I) containing an acidic proton may also beconverted into their non-toxic metal or amine addition salt forms bytreatment with appropriate organic and inorganic bases. Appropriate basesalt forms comprise, for example, the ammonium salts, the alkali andearth alkaline metal salts, e.g. the lithium, sodium, potassium,magnesium, calcium salts and the like, salts with organic bases, e.g.the benzathine, N-methyl-D-glucamine, hydrabamine salts, and salts withamino acids such as, for example, arginine, lysine and the like.

The term addition salt as used hereinabove also comprises the solvateswhich the compounds of formula (I) as well as the salts thereof, areable to form. Such solvates are for example hydrates, alcoholates andthe like.

The term “quaternary amine” as used hereinbefore defines the quaternaryammonium salts which the compounds of formula (I) are able to form byreaction between a basic nitrogen of a compound of formula (I) and anappropriate quaternizing agent, such as, for example, an optionallysubstituted alkylhalide, arylhalide or arylalkylhalide, e.g.methyliodide or benzyliodide. Other reactants with good leaving groupsmay also be used, such as alkyl trifluoromethanesulfonates, alkylmethanesulfonates, and alkyl p-toluenesulfonates. A quaternary amine hasa positively charged nitrogen. Pharmaceutically acceptable counterionsinclude chloro, bromo, iodo, trifluoroacetate and acetate. Thecounterion of choice can be introduced using ion exchange resins.

The N-oxide forms of the present compounds are meant to comprise thecompounds of formula (I) wherein one or several nitrogen atoms areoxidized to the so-called N-oxide.

It will be appreciated that the compounds of formula (I) may have metalbinding, chelating, complex forming properties and therefore may existas metal complexes or metal chelates. Such metalated derivatives of thecompounds of formula (I) are intended to be included within the scope ofthe present invention.

Some of the compounds of formula (I) may also exist in their tautomericform. Such forms although not explicitly indicated in the above formulaare intended to be included within the scope of the present invention.

As mentioned above, the compounds of formula (I) have several asymmetriccenters. In order to more efficiently refer to each of these asymmetriccenters, the numbering system as indicated in the following structuralformula will be used.

Asymmetric centers are present at positions 1, 4 and 6 of the macrocycleas well as at the carbon atom 3′ in the 5-membered ring, carbon atom 2′when the R² substituent is C₁₋₆alkyl, and at carbon atom 1′ when X isCH. Each of these asymmetric centers can occur in their R or Sconfiguration.

The stereochemistry at position 1 preferably corresponds to that of anL-amino acid configuration, i.e. that of L-proline.

When X is CH, the 2 carbonyl groups substituted at positions 1′ and 5′of the cyclopentane ring preferably are in a trans configuration. Thecarbonyl substituent at position 5′ preferably is in that configurationthat corresponds to an L-proline configuration. The carbonyl groupssubstituted at positions 1′ and 5′ preferably are as depicted below inthe structure of the following formula

The compounds of formula (I) include a cyclopropyl group as representedin the structural fragment below:

wherein C₇ represents the carbon at position 7 and carbons at position 4and 6 are asymmetric carbon atoms of the cyclopropane ring.

Notwithstanding other possible asymmetric centers at other segments ofthe compounds of formula (I), the presence of these two asymmetriccenters means that the compounds can exist as mixtures of diastereomers,such as the diastereomers of compounds of formula (I) wherein the carbonat position 7 is configured either syn to the carbonyl or syn to theamide as shown below.

One embodiment concerns compounds of formula (I) wherein the carbon atposition 7 is configured syn to the carbonyl. Another embodimentconcerns compounds of formula (I) wherein the configuration at thecarbon at position 4 is R. A specific subgroup of compounds of formula(I) are those wherein the carbon at position 7 is configured syn to thecarbonyl and wherein the configuration at the carbon at position 4 is R.

The compounds of formula (I) may include as well a proline residue (whenX is N) or a cyclopentyl or cyclopentenyl residue (when X is CH or C).Preferred are the compounds of formula (I) wherein the substituent atthe 1 (or 5′) position and the substituent at position 3′ are in a transconfiguration. Of particular interest are the compounds of formula (I)wherein position 1 has the configuration corresponding to L-proline andthe substituent at position 3′ is in a trans configuration in respect ofposition 1. Preferably the compounds of formula (I) have thestereochemistry as indicated in the structures of formulae (I-a) and(I-b) below:

One embodiment of the present invention concerns compounds of formula(I) or of formula (I-a) or of any subgroup of compounds of formula (I),wherein one or more of the following conditions apply:

-   (a) R² is hydrogen;-   (b) X is nitrogen;-   (c) a double bond is present between carbon atoms 7 and 8.

One embodiment of the present invention concerns compounds of formula(I) or of formulae (I-a), (I-b), or of any subgroup of compounds offormula (I), wherein one or more of the following conditions apply:

-   (a) R² is hydrogen;-   (b) X is CH;-   (c) a double bond is present between carbon atoms 7 and 8.

Particular subgroups of compounds of formula (I) are those representedby the following structural formulae:

Amongst the compounds of formula (I-c) and (I-d), those having thestereochemical configuration of the compounds of formulae (I-a), and(I-b), respectively, are of particular interest.

The double bond between carbon atoms 7 and 8 in the compounds of formula(I), or in any subgroup of compounds of formula (I), may be in a cis orin a trans configuration. Preferably the double bond between carbonatoms 7 and 8 is in a cis configuration, as depicted in formulae (I-c)and (I-d).

A double bond between carbon atoms 1′ and 2′ may be present in thecompounds of formula (I), or in any subgroup of compounds of formula(I), as depicted in formula (I-e) below.

Yet another particular subgroup of compounds of formula (I) are thoserepresented by the following structural formulae:

Amongst the compounds of formulae (I-f), (I-g) or (I-h), those havingthe stereochemical configuration of the compounds of formulae (I-a) and(I-b) are of particular interest.

In (I-a), (I-b), (I-c), (I-d), (I-e), (I-f), (I-g) and (I-h), whereapplicable, X, n, R¹, R², R³, R⁴, R⁵, and R⁶ are as specified in thedefinitions of the compounds of formula (I) or in any of the subgroupsof compounds of formula (I) specified herein.

It is to be understood that the above defined subgroups of compounds offormulae (I-a), (I-b), (I-c), (I-d), (I-e), (I-f), (I-g) or (I-h), aswell as any other subgroup defined herein, are meant to also compriseany N-oxides, addition salts, quaternary amines, metal complexes andstereochemically isomeric forms of such compounds.

When n is 2, the moiety —CH₂— bracketed by “n” corresponds to ethanediylin the compounds of formula (I) or in any subgroup of compounds offormula (I). When n is 3, the moiety —CH₂— bracketed by “n” correspondsto propanediyl in the compounds of formula (I) or in any subgroup ofcompounds of formula (I). When n is 4, the moiety —CH₂— bracketed by “n”corresponds to butanediyl in the compounds of formula (I) or in anysubgroup of compounds of formula (I). When n is 5, the moiety—CH₂-bracketed by “n” corresponds to pentanediyl in the compounds offormula (I) or in any subgroup of compounds of formula (I). When n is 6,the moiety —CH₂— bracketed by “n” corresponds to hexanediyl in thecompounds of formula (I) or in any subgroup of compounds of formula (I).Particular subgroups of the compounds of formula (I) are those compoundswherein n is 4 or 5.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein

-   (a) R¹ is —OR⁷, in particular wherein R⁷ is C₁₋₆alkyl, such as    methyl, ethyl, or tert-butyl (or t.butyl) and most preferably where    R⁷ is hydrogen;-   (b) R¹ is —NHS(═O)₂R⁸, in particular wherein R⁸ is C₁₋₆alkyl,    C₃-C₇cycloalkyl, or aryl, e.g. wherein R⁸ is methyl, cyclopropyl, or    phenyl; or-   (c) R¹ is —NHS(═O)₂R⁸, in particular wherein R⁸ is C₃₋₇cycloalkyl    substituted with C₁₋₆alkyl, preferably wherein R⁸ is cyclopropyl,    cyclobutyl, cyclopentyl, or cyclohexyl, any of which is substituted    with C₁₋₄alkyl, i.e. with methyl, ethyl, propyl, isopropyl, butyl,    tert-butyl, or isobutyl.

Further embodiments of the invention are compounds of formula (I) or anyof the subgroups of compounds of formula (I) wherein R¹ is —NHS(═O)₂R⁸,in particular wherein R⁸ is cyclopropyl substituted with C₁₋₄alkyl, i.e.with methyl, ethyl, propyl, or isopropyl.

Further embodiments of the invention are compounds of formula (I) or anyof the subgroups of compounds of formula (I) wherein R¹ is —NHS(═O)₂R⁸,in particular wherein R⁸ is 1-methylcyclopropyl.

Further embodiments of the invention are compounds of formula (I) or anyof the subgroups of compounds of formula (I) wherein

-   (a) R² is hydrogen;-   (b) R² is C₁₋₆alkyl, preferably methyl.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein

-   (a) X is N, C (X being linked via a double bond) or CH (X being    linked via a single bond) and R² is hydrogen;-   (b) X is C (X being linked via a double bond) and R² is C₁₋₆alkyl,    preferably methyl.

Further embodiments of the invention are compounds of formula (I) or anyof the subgroups of compounds of formula (I) wherein

-   (a) R³ is hydrogen;-   (b) R³ is C₁₋₆alkyl;-   (c) R³ is C₁₋₆alkoxyC₁₋₆alkyl or C₃₋₇cycloalkyl.

Preferred embodiments of the invention are compounds of formula (I) orany of the subgroups of compounds of formula (I) wherein R³ is hydrogen,or C₁₋₆alkyl, more preferably hydrogen or methyl.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R⁴ is aryl or Het, eachindependently, optionally substituted with any of the substituents ofHet or aryl mentioned in the definitions of the compounds of formula (I)or of any of the subgroups of compounds of formula (I); or specificallysaid aryl or Het being each, independently, optionally substituted withC₁₋₆alkyl, halo, amino, mono- or diC₁₋₆alkylamino, pyrrolidinyl,piperidinyl, morpholinyl, piperazinyl, 4-C₁₋₆alkylpiperazinyl; andwherein the morpholinyl and piperidinyl groups may optionallysubstituted with one or two C₁₋₆alkyl radicals;

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R⁴ is a radical

or, in particular, wherein R⁴ is selected from the group consisting of:

wherein, where possible a nitrogen may bear an R^(4a) substituent or alink to the remainder of the molecule; each R^(4a) in any of the R⁴substituents may be selected from those mentioned as possiblesubstituents on Het, as specified in the definitions of the compounds offormula (I) or of any of the subgroups of compounds of formula (I);

more specifically each R^(4a) may be hydrogen, halo, C₁₋₆alkyl, amino,or mono- or di-C₁₋₆alkylamino, pyrrolidinyl, piperidinyl, morpholinyl,piperazinyl, 4-C₁₋₆alkyl-piperazinyl; and wherein the morpholinyl andpiperidinyl groups may optionally substituted with one or two C₁₋₆alkylradicals;

more specifically each R^(4a) is, each independently, hydrogen, halo,C₁₋₆alkyl, amino, or mono- or di-C₁₋₆alkylamino;

and where R^(4a) is substituted on a nitrogen atom, it preferably is acarbon containing substituent that is connected to the nitrogen via acarbon atom or one of its carbon atoms; and wherein in that instanceR^(4a) preferably is C₁₋₆alkyl.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R⁴ is phenyl or pyridiyl(in particular 4-pyridyl) which each may be substituted with 1, 2 or 3substituents selected from those mentioned for aryl in the definitionsof the compounds of formula (I) or of any of the subgroups thereof. Inparticular said phenyl or pyridyl is substituted with 1-3 (or with 1-2,or with one) substituent or substituents selected from halo, C₁₋₆alkylor C₁₋₆alkoxy.

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R⁵ is halo, or C₁₋₆alkyl,preferably methyl, ethyl, isopropyl, tert-butyl, fluoro, chloro, orbromo. include poluhaloC₁₋₆alkyl

Embodiments of the invention are compounds of formula (I) or any of thesubgroups of compounds of formula (I) wherein R⁶ is C₁₋₆alkoxy ordiC₁₋₆alkylamino; preferably R⁶ is methoxy or dimethylamino; morepreferably R⁶ is methoxy.

The compounds of formula (I) consist of three building blocks P1, P2,P3. Building block P1 further contains a P1′ tail. The carbonyl groupmarked with an asterisk in compound (I-c) below may be part of eitherbuilding block P2 or of building block P3. For reasons of chemistry,building block P2 of the compounds of formula (I) wherein X is Cincorporates the carbonyl group attached to the position 1′.

The linking of building blocks P1 with P2, P2 with P3, and P1 with P1′(when R¹ is —NH—SO₂R⁸ or —OR) involves forming an amide bond. Thelinking of blocks P1 and P3 involves double bond formation. The linkingof building blocks P1, P2 and P3 to prepare compounds (I-i) or (I-j) canbe done in any given sequence. One of the steps involves a cyclizationwhereby the macrocycle is formed.

Represented herebelow are compounds (I-i) which are compounds of formula(I) wherein carbon atoms C7 and C8 are linked by a double bond, andcompounds (I-j) which are compounds of formula (I) wherein carbon atomsC7 and C8 are linked by a single bond. The compounds of formula (I-j)can be prepared from the corresponding compounds of formula (I-I) byreducing the double bond in the macrocycle.

It should be noted that in compounds of formula (I-c), the amide bondformation between blocks P2 and P3 may be accomplished at two differentpositions of the urea fragment. A first amide bond encompasses thenitrogen of the pyrrolidine ring and the adjacent carbonyl (marked withan asterisk). An alternative second amide bond formation involves thereaction of the asterisked carbonyl with a —NHR³ group. Both amide bondformations between building blocks P2 and P3 are feasible.

The synthesis procedures described hereinafter are meant to beapplicable for as well the racemates, stereochemically pureintermediates or end products, or any stereoisomeric mixtures. Theracemates or stereochemical mixtures may be separated intostereoisomeric forms at any stage of the synthesis procedures. In oneembodiment, the intermediates and end products have the stereochemistryspecified above in the compounds of formula (I-a) and (I-b).

In order to simplify the structural representation of the compounds offormula (I) or the intermediates the group

is represented by R⁹ and the dotted line represents the bond linkingsaid group represented by R⁹ to the remainder of the molecule.

In one embodiment, compounds (I-i) are prepared by first forming theamide bonds and subsequent forming the double bond linkage between P3and P1 with concomitant cyclization to the macrocycle.

In a preferred embodiment, compounds (I) wherein the bond between C₇ andC₈ is a double bond, which are compounds of formula (I-i), as definedabove, may be prepared as outlined in the following reaction scheme:

Formation of the macrocycle can be carried out via an olefin metathesisreaction in the presence of a suitable metal catalyst such as e.g. theRu-based catalyst reported by Miller, S. J., Blackwell, H. E., Grubbs,R. H. J. Am. Chem. Soc. 118, (1996), 9606-9614; Kingsbury, J. S.,Harrity, J. P. A., Bonitatebus, P. J., Hoveyda, A. H., J. Am. Chem. Soc.121, (1999), 791-799; and Huang et al., J. Am. Chem. Soc. 121, (1999),2674-2678; for example a Hoveyda-Grubbs catalyst.

Air-stable ruthenium catalysts such asbis(tricyclohexylphosphine)-3-phenyl-1H-inden-1-ylidene rutheniumchloride (Neolyst M1®) orbis(tricyclohexylphosphine)-[(phenylthio)methylene]ruthenium (IV)dichloride can be used. Other catalysts that can be used are Grubbsfirst and second generation catalysts, i.e.Benzylidene-bis(tricycle-hexylphosphine)dichlororuthenium and(1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(phenylmethylene)-(tricyclohexylphosphine)ruthenium,respectively. Of particular interest are the Hoveyda-Grubbs first andsecond generation catalysts, which aredichloro(o-isopropoxyphenylmethylene)(tricyclohexylphosphine)-ruthenium(II)and1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro-(o-isopropoxyphenylmethylene)rutheniumrespectively. Also other catalysts containing other transition metalssuch as Mo can be used for this reaction.

The metathesis reactions may be conducted in a suitable solvent such asfor example ethers, e.g. THF, dioxane; halogenated hydrocarbons, e.g.dichoromethane, CHCl₃, 1,2-dichloroethane and the like, hydrocarbons,e.g. toluene. In a preferred embodiment, the metathesis reaction isconducted in toluene. These reactions are conducted at increasedtemperatures under nitrogen atmosphere.

Compounds of formula (I) wherein the link between C7 and C8 in themacrocycle is a single bond, i.e. compounds of formula (I-j), can beprepared from the compounds of formula (I-i) by a reduction of the C7-C8double bond in the compounds of formula (I-i). This reduction may beconducted by catalytic hydrogenation with hydrogen in the presence of anoble metal catalyst such as, for example, Pt, Pd, Rh, Ru or Raneynickel. Of interest is Rh on alumina. The hydrogenation reactionpreferably is conducted in a solvent such as, e.g. an alcohol such asmethanol, ethanol, or an ether such as THF, or mixtures thereof. Watercan also be added to these solvents or solvent mixtures.

The R¹ group can be connected to the P1 building block at any stage ofthe synthesis, i.e. before or after the cyclization, or before or afterthe cyclization and reduction as described herein above. The compoundsof formula (I) wherein R¹ represents —NHSO₂R⁸, said compounds beingrepresented by formula (I-k-1), can be prepared by linking the R¹ groupto P1 by forming an amide bond between both moieties. Similarly, thecompounds of formula (I) wherein R¹ represents —OR⁷, i.e. compounds(I-k-2), can be prepared by linking the R¹ group to P1 by forming anester bond. In one embodiment, the —OR⁵ groups are introduced in thelast step of the synthesis of the compounds (I) as outlined in thefollowing reaction schemes wherein G represents a group:

Intermediate (2a) can be coupled with the amine (2b) by an amide formingreaction such as any of the procedures for the formation of an amidebond described hereinafter. In particular, (2a) may be treated with acoupling agent, for example N,N′-carbonyl-diimidazole (CDI), EEDQ, IIDQ,EDCI or benzotriazol-1-yl-oxy-tris-pyrrolidino-phosphoniumhexafluorophosphate (commercially available as PyBOP®), in a solventsuch as an ether, e.g. THF, or a halogenated hydrocarbon, e.g.dichloromethane, chlorophorm, dichloroethane, and reacted with thedesired sulfonamide (2b), preferably after reacting (2a) with thecoupling agent. The reactions of (2a) with (2b) preferably are conductedin the presence of a base, for example a trialkylamine such astriethylamine or diisopropylethylamine, or1,8-diazabicycle[5.4.0]undec-7-ene (DBU). Intermediate (2a) can also beconverted into an activated form, e.g. an activated form of generalformula G-CO—Z, wherein Z represents halo, or the rest of an activeester, e.g. Z is an aryloxy group such as phenoxy, p.nitrophenoxy,pentafluorophenoxy, trichlorophenoxy, pentachlorophenoxy and the like;or Z can be the rest of a mixed anhydride. In one embodiment, G-CO—Z isan acid chloride (G-CO—CI) or a mixed acid anhydride (G-CO—O—CO—R orG-CO—O—CO—OR, R in the latter being e.g. C₁₋₄alkyl, such as methyl,ethyl, propyl, i.propyl, butyl, t.butyl, i.butyl, or benzyl). Theactivated form G-CO—Z is reacted with the sulfonamide (2b).

The activation of the carboxylic acid in (2a) as described in the abovereactions may lead to an internal cyclization reaction to an azalactoneintermediate of formula

wherein X, R², R³, R⁹, n are as specified above and wherein thestereogenic centers may have the stereochemical configuration asspecified above, for example as in (I-a) or (I-b). The intermediates(2a-1) can be isolated from the reaction mixture, using conventionalmethodology, and the isolated intermediate (2a-1) is then reacted with(2b), or the reaction mixture containing (2a-1) can be reacted furtherwith (2b) without isolation of (2a-1). In one embodiment, where thereaction with the coupling agent is conducted in a water-immisciblesolvent, the reaction mixture containing (2a-1) may be washed with wateror with slightly basic water in order to remove all water-soluble sideproducts. The thus obtained washed solution may then be reacted with(2b) without additional purification steps. The isolation ofintermediates (2a-1) on the other hand may provide certain advantages inthat the isolated product, after optional further purification, may bereacted with (2b), giving rise to less side products and an easierwork-up of the reaction.

Intermediate (2a) can be coupled with the alcohol (2c) by an esterforming reaction. For example, (2a) and (2c) are reacted together withremoval of water either physically, e.g. by azeotropical water removal,or chemically by using a dehydrating agent. Intermediate (2a) can alsobe converted into an activated form G-CO—Z, such as the activated formsmentioned above, and subsequently reacted with the alcohol (2c). Theester forming reactions preferably are conducted in the presence of abase such as an alkali metal carbonate or hydrogen carbonate, e.g.sodium or potassium hydrogen carbonate, or a tertiary amine such as theamines mentioned herein in relation to the amide forming reactions, inparticular a trialkylamine, e.g. triethylamine Solvents that can be usedin the ester forming reactions comprise ethers such as THF; halogenatedhydrocarbons such as dichoromethane, CH₂Cl₂; hydrocarbons such astoluene; polar aprotic solvents such as DMF, DMSO, DMA; and the likesolvents.

The compounds of formula (I) wherein R³ is hydrogen, said compoundsbeing represented by (I-1), can also be prepared by removal of aprotecting group PG, from a corresponding nitrogen-protectedintermediate (3a), as in the following reaction scheme. The protectinggroup PG in particular is any of the nitrogen protecting groupsmentioned hereinafter and can be removed using procedures also mentionedhereinafter:

The starting materials (3a) in the above reaction can be preparedfollowing the procedures for the preparation of compounds of formula(I), but using intermediates wherein the group R³ is PG.

The compounds of formula (I) can also be prepared by reacting anintermediate (4a) with intermediate (4b) as outlined in the followingreaction scheme wherein the various radicals have the meanings specifiedabove:

Y in (4b) represents hydroxy or a leaving group LG such as a halide,e.g. bromide or chloride, or an arylsulfonyl group, e.g. mesylate,triflate or tosylate and the like.

In one embodiment, the reaction of (4a) with (4b) is an O-arylationreaction and Y represents a leaving group. This reaction can beconducted following the procedures described by E. M. Smith et al. (J.Med. Chem. (1988), 31, 875-885). In particular, this reaction isconducted in the presence of a base, preferably a strong base, in areaction-inert solvent, e.g. one of the solvents mentioned for theformation of an amide bond.

In a particular embodiment, starting material (4a) is reacted with (4b)in the presence of a base which is strong enough to detract a hydrogenfrom the hydroxy group, for example an alkali of alkaline metal hydridesuch as LiH or sodium hydride, or alkali metal alkoxide such as sodiumor potassium methoxide or ethoxide, potassium tert-butoxide, in areaction inert solvent like a dipolar aprotic solvent, e.g. DMA, DMF andthe like. The resulting alcoholate is reacted with the arylating agent(4b), wherein Y is a suitable leaving group as mentioned above. Theconversion of (4a) to (I) using this type of O-arylation reaction doesnot change the stereochemical configuration at the carbon bearing thehydroxy group.

Alternatively, the reaction of (4a) with (4b) can also be conducted viaa Mitsunobu reaction (Mitsunobu, 1981, Synthesis, January, 1-28; Rano etal., Tetrahedron Lett., 1995, 36, 22, 3779-3792; Krchnak et al.,Tetrahedron Lett., 1995, 36, 5, 6193-6196; Richter et al., TetrahedronLett., 1994, 35, 27, 4705-4706). This reaction comprises treatment ofintermediate (4a) with (4b) wherein Y is hydroxyl, in the presence oftriphenylphosphine and an activating agent such as a dialkylazocarboxylate, e.g. diethyl azodicarboxylate (DEAD), diisopropylazodicarboxylate (DIAD) or the like. The Mitsunobu reaction changes thestereochemical configuration at the carbon bearing the hydroxy group.

Alternatively, in order to prepare the compounds of formula (I), firstan amide bond between building blocks P2 and P1 is formed, followed bycoupling of the P3 building block to the P1 moiety in P1-P2, and asubsequent carbamate or ester bond formation between P3 and the P2moiety in P2-P1-P3 with concomitant ring closure.

Yet another alternative synthetic methodology is the formation of anamide bond between building blocks P2 and P3, followed by the couplingof building block P1 to the P3 moiety in P3-P2, and a last amide bondformation between P1 and P2 in P1-P3-P2 with concomitant ring closure.

Building blocks P1 and P3 can be linked to a P1-P3 sequence. If desired,the double bond linking P1 and P3 may be reduced. The thus formed P1-P3sequence, either reduced or not, can be coupled to building block P2 andthe thus forming sequence P1-P3-P2 subsequently cyclized, by forming anamide bond.

Building blocks P1 and P3 in any of the previous approaches can belinked via double bond formation, e.g. by the olefin metathesis reactiondescribed hereinafter, or a Wittig type reaction. If desired, the thusformed double bond can be reduced, similarly as described above for theconversion of (I-i) to (I-j). The double bond can also be reduced at alater stage, i.e. after addition of a third building block, or afterformation of the macrocycle. Building blocks P2 and P1 are linked byamide bond formation and P3 and P2 are linked by carbamate or esterformation.

The tail P1′ can be bonded to the P1 building block at any stage of thesynthesis of the compounds of formula (I), for example before or aftercoupling the building blocks P2 and P1; before or after coupling the P3building block to P1; or before or after ring closure.

The individual building blocks can first be prepared and subsequentlycoupled together or alternatively, precursors of the building blocks canbe coupled together and modified at a later stage to the desiredmolecular composition.

The functionalities in each of the building blocks may be protected toavoid side reactions.

The formation of amide bonds can be carried out using standardprocedures such as those used for coupling amino acids in peptidesynthesis. The latter involves the dehydrative coupling of a carboxylgroup of one reactant with an amino group of the other reactant to forma linking amide bond. The amide bond formation may be performed byreacting the starting materials in the presence of a coupling agent orby converting the carboxyl functionality into an active form such as anactive ester, mixed anhydride or a carboxyl acid chloride or bromide.General descriptions of such coupling reactions and the reagents usedtherein can be found in general textbooks on peptide chemistry, forexample, M. Bodanszky, “Peptide Chemistry”, 2nd rev. ed.,Springer-Verlag, Berlin, Germany, (1993).

Examples of coupling reactions with amide bond formation include theazide method, mixed carbonic-carboxylic acid anhydride (isobutylchloroformate) method, the carbodiimide (dicyclohexylcarbodiimide,diisopropylcarbodiimide, or water-soluble carbodiimide such asN-ethyl-N′-[(3-dimethylamino)propyl]carbodiimide) method, the activeester method (e.g. p-nitrophenyl, p-chlorophenyl, trichlorophenyl,pentachloro-phenyl, pentafluorophenyl, N-hydroxysuccinic imido and thelike esters), the Woodward reagent K-method, the 1,1-carbonyldiimidazole(CDI or N,N′-carbonyl-diimidazole) method, the phosphorus reagents oroxidation-reduction methods. Some of these methods can be enhanced byadding suitable catalysts, e.g. in the carbodiimide method by adding1-hydroxybenzotriazole, DBU (1,8-diazabicyclo[5.4.0]undec-7-ene), or4-DMAP. Further coupling agents are(benzotriazol-1-yloxy)tris-(dimethylamino) phosphoniumhexafluorophosphate, either by itself or in the presence of1-hydroxy-benzotriazole or 4-DMAP; or2-(1H-benzotriazol-1-yl)-N,N,N′,N′-tetra-methyluroniumtetrafluoroborate, orO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate. These coupling reactions can be performed in eithersolution (liquid phase) or solid phase.

A preferred amide bond formation is performed employingN-ethyloxycarbonyl-2-ethyloxy-1,2-dihydroquinoline (EEDQ) orN-isobutyloxy-carbonyl-2-isobutyloxy-1,2-dihydroquinoline (IIDQ). Unlikethe classical anhydride procedure, EEDQ and IIDQ do not require base norlow reaction temperatures. Typically, the procedure involves reactingequimolar amounts of the carboxyl and amine components in an organicsolvent (a wide variety of solvents can be used). Then EEDQ or IIDQ isadded in excess and the mixture is allowed to stir at room temperature.

The coupling reactions preferably are conducted in an inert solvent,such as halogenated hydrocarbons, e.g. dichloromethane, chloroform,dipolar aprotic solvents such as acetonitrile, dimethylformamide,dimethylacetamide, DMSO, HMPT, ethers such as tetrahydrofuran (THF).

In many instances the coupling reactions are done in the presence of asuitable base such as a tertiary amine, e.g. triethylamine,diisopropylethylamine (DIPEA), N-methyl-morpholine, N-methylpyrrolidine,4-DMAP or 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). The reactiontemperature may range between 0° C. and 50° C. and the reaction time mayrange between 15 min and 24 h.

The functional groups in the building blocks that are linked togethermay be protected to avoid formation of undesired bonds. Appropriateprotecting groups that can be used are listed for example in Greene,“Protective Groups in Organic Chemistry”, John Wiley & Sons, New York(1999) and “The Peptides: Analysis, Synthesis, Biology”, Vol. 3,Academic Press, New York (1987).

Carboxyl groups can be protected as an ester that can be cleaved off togive the carboxylic acid. Protecting groups that can be used include 1)alkyl esters such as methyl, trimethylsilyl and tert-butyl; 2) arylalkylesters such as benzyl and substituted benzyl; or 3) esters that can becleaved by a mild base or mild reductive means such as trichloroethyland phenacyl esters.

Amino groups can be protected by a variety of N-protecting groups, suchas:

-   1) acyl groups such as formyl, trifluoroacetyl, phthalyl, and    p-toluenesulfonyl;-   2) aromatic carbamate groups such as benzyloxycarbonyl (Cbz or Z)    and substituted benzyloxycarbonyls, and 9-fluorenylmethyloxycarbonyl    (Fmoc);-   3) aliphatic carbamate groups such as tert-butyloxycarbonyl (Boc),    ethoxycarbonyl, diisopropylmethoxy-carbonyl, and allyloxycarbonyl;-   4) cyclic alkyl carbamate groups such as cyclopentyloxycarbonyl and    adamantyloxycarbonyl;-   5) alkyl groups such as triphenylmethyl, benzyl or substituted    benzyl such as 4-methoxybenzyl;-   6) trialkylsilyl such as trimethylsilyl or t.Bu dimethylsilyl; and-   7) thiol containing groups such as phenylthiocarbonyl and    dithiasuccinoyl. Interesting amino protecting groups are Boc and    Fmoc.

Preferably the amino protecting group is cleaved off prior to the nextcoupling step. Removal of N-protecting groups can be done followingart-known procedures. When the Boc group is used, the methods of choiceare trifluoroacetic acid, neat or in dichloromethane, or HCl in dioxaneor in ethyl acetate. The resulting ammonium salt is then neutralizedeither prior to the coupling or in situ with basic solutions such asaqueous buffers, or tertiary amines in dichloromethane or acetonitrileor dimethyl-formamide. When the Fmoc group is used, the reagents ofchoice are piperidine or substituted piperidine in dimethylformamide,but any secondary amine can be used. The deprotection is carried out ata temperature between 0° C. and room temperature, usually around 15-25°C., or 20-22° C.

Other functional groups that can interfere in the coupling reactions ofthe building blocks may also be protected. For example hydroxyl groupsmay be protected as benzyl or substituted benzyl ethers, e.g.4-methoxybenzyl ether, benzoyl or substituted benzoyl esters, e.g.4-nitrobenzoyl ester, or with trialkylsilyl groups (e.g. trimethylsilylor tert-butyldimethylsilyl).

Further amino groups may be protected by protecting groups that can becleaved off selectively. For example, when Boc is used as the α-aminoprotecting group, the following side chain protecting groups aresuitable: p-toluenesulfonyl (tosyl) moieties can be used to protectfurther amino groups; benzyl (Bn) ethers can be used to protect hydroxygroups; and benzyl esters can be used to protect further carboxylgroups. Or when Fmoc is chosen for the α-amino protection, usuallytert-butyl based protecting groups are acceptable. For instance, Boc canbe used for further amino groups; tert-butyl ethers for hydroxyl groups;and tert-butyl esters for further carboxyl groups.

Any of the protecting groups may be removed at any stage of thesynthesis procedure but preferably, the protecting groups of any of thefunctionalities not involved in the reaction steps are removed aftercompletion of the build-up of the macrocycle. Removal of the protectinggroups can be done in whatever manner is dictated by the choice ofprotecting groups, which manners are well known to those skilled in theart.

The intermediates of formula (1a) wherein X is N, said intermediatesbeing represented by formula (1a-1), may be prepared starting fromintermediates (5a) which are reacted with an alkenamine (5b) in thepresence of a carbonyl introducing agent as outlined in the followingreaction scheme.

Carbonyl (CO) introducing agents include phosgene, or phosgenederivatives such as carbonyl diimidazole (CDI), and the like. In oneembodiment (5a) is reacted with the CO introducing agent in the presenceof a suitable base and a solvent, which can be the bases and solventsused in the amide forming reactions as described above. In a particularembodiment, the base is a hydrogencarbonate, e.g. NaHCO₃, or a tertiaryamine such as triethylamine and the like, and the solvent is an ether orhalogenated hydrocarbon, e.g. THF, CH₂Cl₂, CHCl₃, and the like.Thereafter, the amine (5b) is added thereby obtaining intermediates(1a-1) as in the above scheme. An alternative route using similarreaction conditions involves first reacting the CO introducing agentwith the alkenamine (5b) and then reacting the thus formed intermediatewith (5a).

The intermediates (1a-1) can alternatively be prepared as follows:

PG¹ is an O-protecting group, which can be any of the groups mentionedherein and in particular is a benzoyl or substituted benzoyl group suchas 4-nitrobenzoyl. In the latter instance this group can be removed byreaction with a an alkali metal hydroxide (LiOH, NaOH, KOH), inparticular where PG¹ is 4-nitrobenzoyl, with LiOH, in an aqueous mediumcomprising water and a water-soluble organic solvent such as an alkanol(methanol, ethanol) and THF.

Intermediates (6a) are reacted with (5b) in the presence of a carbonylintroducing agent, similar as described above, and this reaction yieldsintermediates (6c). These are deprotected, in particular using thereaction conditions mentioned above. The resulting alcohol (6d) isreacted with intermediates (4b) as described above for the reaction of(4a) with (4b) and this reaction results in intermediates (1a-1).

The intermediates of formula (1a) wherein X is C, said intermediatesbeing represented by formula (1a-2), may be prepared by an amide formingreaction starting from intermediates (7a) which are reacted with anamine (5b) as shown in the following reaction scheme, using reactionconditions for preparing amides such as those described above.

The intermediates (1a-1) can alternatively be prepared as follows:

PG¹ is an O-protecting group as described above. The same reactionconditions as described above may be used: amide formation as describedabove, removal of PG¹ as in the description of the protecting groups andintroduction of R⁹ as in the reactions of (4a) with the reagents (4b).

The intermediates of formula (2a) may be prepared by first cyclizing theopen amide (9a) to a macrocyclic ester (9b), which in turn is convertedto (2a) as follows:

PG² is a carboxyl protecting group, e.g. one of the carboxyl protectinggroups mentioned above, in particular a C₁₋₄alkyl or benzyl ester, e.g.a methyl, ethyl or t.butyl ester. The reaction of (9a) to (9b) is ametathesis reaction and is conducted as described above. The group PG²is removed following procedures also described above. Where PG¹ is aC₁₋₄alkyl ester, it is removed by alkaline hydrolysis, e.g. with NaOH orpreferably LiOH, in an aqueous solvent, e.g. a C₁₋₄alkanol/watermixture. A benzyl group can be removed by catalytic hydrogenation.

In an alternative synthesis, intermediates (2a) can be prepared asfollows:

The PG¹ group is selected such that it is selectively cleavable towardsPG². PG² may be e.g. methyl or ethyl esters, which can be removed bytreatment with an alkali metal hydroxide in an aqueous medium, in whichcase PG¹ e.g. is t.butyl or benzyl. PG² may be t.butyl esters removableunder weakly acidic conditions or PG¹ may be benzyl esters removablewith strong acid or by catalytic hydrogenation, in the latter two casesPG¹ e.g. is a benzoic ester such as a 4-nitrobenzoic ester.

First, intermediates (10a) are cyclized to the macrocyclic esters (10b),the latter are deprotected by removal of the PG¹ group to (10c), whichare reacted with intermediates (4b), followed by removal of carboxylprotecting group PG². The cyclization, deprotection of PG¹ and PG² andthe coupling with (4b) are as described above.

The R¹ groups can be introduced at any stage of the synthesis, either asthe last step as described above, or earlier, before the macrocycleformation. In the following scheme, the groups R¹ being —NH—SO₂R⁸ or—OR⁷ (which are as specified above) are introduced:

In the above scheme, PG² is as defined above and L¹ is a P3 group

wherein n and R³ are as defined above and where X is N, L¹ may also be anitrogen-protecting group (PG, as defined above) and where X is C, L¹may also be a group —COOPG^(2a), wherein the group PG^(2a) is a carboxylprotecting group similar as PG², but wherein PG^(2a) is selectivelycleavable towards PG². In one embodiment PG^(2a) is t.butyl and PG² ismethyl or ethyl.

The intermediates (11c) and (11d) wherein L¹ represents a group (b)correspond to the intermediates (1a) and may be processed further asspecified above.

Coupling of P1 and P2 Building Blocks

The P1 and P2 building blocks are linked using an amide forming reactionfollowing the procedures described above. The P1 building block may havea carboxyl protecting group PG² (as in (12b)) or may already be linkedto P1′ group (as in (12c)). L² is a N-protecting group (PG), or a group(b), as specified above. L³ is hydroxy, —OPG¹ or a group —O—R⁹ asspecified above. Where in any of the following reaction schemes L³ ishydroxy, prior to each reaction step, it may be protected as a group—OPG¹ and, if desired, subsequently deprotected back to a free hydroxyfunction. Similarly as described above, the hydroxy function may beconverted to a group —O—R⁹.

In the procedure of the above scheme, a cyclopropyl amino acid (12b) or(12c) is coupled to the acid function of the P2 building block (12a)with the formation of an amide linkage, following the proceduresdescribed above. Intermediates (12d) or (12e) are obtained. Where in thelatter L² is a group (b), the resulting products are P3-P2-P1 sequencesencompassing some of the intermediates (11c) or (11d) in the previousreaction scheme. Removal of the acid protecting group in (12d), usingthe appropriate conditions for the protecting group used, followed bycoupling with an amine H₂N—SO₂R⁸ (2b) or with HOR⁷ (2c) as describedabove, again yields the intermediates (12e), wherein —COR¹ are amide orester groups. Where L² is a N-protecting group, it can be removedyielding intermediates (5a) or (6a). In one embodiment, PG in thisreaction is a BOC group and PG² is methyl or ethyl. Where additionallyL³ is hydroxy, the starting material (12a) is Boc-L-hydroxyproline. In aparticular embodiment, PG is BOC, PG² is methyl or ethyl and L³ is—O—R⁹.

In one embodiment, L² is a group (b) and these reactions involvecoupling P1 to P2-P3, which results in the intermediates (1a-1) or (1a)mentioned above. In another embodiment, L² is a N-protecting group PG,which is as specified above, and the coupling reaction results inintermediates (12d-1) or (12e-1), from which the group PG can beremoved, using reaction conditions mentioned above, obtainingintermediates (12-f) or respectively (12 g), which encompassintermediates (5a) and (6a) as specified above:

In one embodiment, the group L³ in the above schemes represents a group—O-PG¹ which can be introduced on a starting material (12a) wherein L³is hydroxy. In this instance PG¹ is chosen such that it is selectivelycleavable towards group L² being PG.

In a similar way, P2 building blocks wherein X is C, which arecyclopentane or cyclopentene derivatives, can be linked to P1 buildingblocks as outlined in the following scheme wherein R¹, R², L³ are asspecified above and PG² and PG^(2a) are carboxyl protecting groups.PG^(2a) typically is chosen such that it is selectively cleavabletowards group PG². Removal of the PG^(2a) group in (13c) yieldsintermediates (7a) or (8a), which can be reacted with (5b) as describedabove.

In a particular embodiment, where X is C, R² is H, and where X and thecarbon bearing R² are linked by a single bond (P2 being a cyclopentanemoiety), PG^(2a) and L³ taken together form a bond and the P2 buildingblock is represented by formula:

Bicyclic acid (14a) is reacted with (12b) or (12c) similar as describedabove to (14b) and (14c) respectively, wherein the lactone is openedgiving intermediates (14c) and (14e). The lactones can be opened usingester hydrolysis procedures, for example using the reaction conditionsdescribed above for the alkaline removal of a PG¹ group in (9b), inparticular using basic conditions such as an alkali metal hydroxide,e.g. NaOH, KOH, in particular LiOH.

Intermediates (14c) and (14e) can be processed further as describedhereinafter.

Coupling of P3 and P2 Building Blocks

For P2 building blocks that have a pyrrolidine moiety, the P3 and P2 orP3 and P2-P1 building blocks are linked using a carbamate formingreaction following the procedures described above for the coupling of(5a) with (5b). A general procedure for coupling P2 blocks having apyrrolidine moiety is represented in the following reaction schemewherein L³ is as specified above and L⁴ is a group —O-PG², a group

or a group

In one embodiment L⁴ in (15a) is a group —OPG², the PG² group may beremoved and the resulting acid coupled with cyclopropyl amino acids(12a) or (12b), yielding intermediates (12d) or (12e) wherein L² is aradical (d) or (e).

A general procedure for coupling P3 blocks with a P2 block or a with aP2-P1 block wherein the P2 is a cyclopentane or cyclopentene is shown inthe following scheme. L³ and L⁴ are as specified above.

In a particular embodiment L³ and L⁴ taken together may form a lactonebridge as in (14a), and the coupling of a P3 block with a P2 block is asfollows:

Bicyclic lactone (14a) is reacted with (5b) in an amide forming reactionto amide (16c) in which the lactone bridge is opened to (16d). Thereaction conditions for the amide forming and lactone opening reactionsare as described above or hereinafter. Intermediate (16d) in turn can becoupled to a P1 group as described above.

The reactions in the above schemes are conducted using the sameprocedures as described above for the reactions of (5a), (7a) or (8a)with (5b) and in particular the above reactions wherein L⁴ is a group(d) or (e) correspond to the reactions of (5a), (7a) or (8a) with (5b),as described above.

The building blocks P1, P1′, P2 and P3 used in the preparation of thecompounds of formula (I) can be prepared starting from art-knownintermediates. A number of such syntheses are described hereafter inmore detail.

The individual building blocks can first be prepared and subsequentlycoupled together or alternatively, precursors of the building blocks canbe coupled together and modified at a later stage to the desiredmolecular composition.

The functionalities in each of the building blocks may be protected toavoid side reactions.

Synthesis of P2 Building Blocks

The P2 building blocks contain either a pyrrolidine, a cyclopentane, ora cyclopentene moiety substituted with a group —O—R⁴.

P2 building blocks containing a pyrrolidine moiety can be derived fromcommercially available hydroxy proline.

The preparation of P2 building blocks that contain a cylopentane ringmay be performed as shown in the scheme below.

The bicyclic acid (17b) can be prepared, for example, from3,4-bis(methoxycarbonyl)-cyclopentanone (17a), as described byRosenquist et al. in Acta Chem. Scand. 46 (1992) 1127-1129. A first stepin this procedure involves the reduction of the keto group with areducing agent like sodium borohydride in a solvent such as methanol,followed by hydrolysis of the esters and finally ring closure to thebicyclic lactone (17b) using lactone forming procedures, in particularby using acetic anhydride in the presence of a weak base such aspyridine. The carboxylic acid functionality in (17b) can then beprotected by introducing an appropriate carboxyl protecting group, suchas a group PG², which is as specified above, thus providing bicyclicester (17c). The group PG² in particular is acid-labile such as at.butyl group and is introduced e.g. by treatment with isobutene in thepresence of a Lewis acid or with di-tert-butyl dicarbonate in thepresence of a base such as a tertiary amine like dimethylamino-pyridineor triethylamine in a solvent like dichloromethane. Lactone opening of(17c) using reaction conditions described above, in particular withlithium hydroxide, yields the acid (17d), which can be used further incoupling reactions with P1 building blocks. The free acid in (17d) mayalso be protected, preferably with an acid protecting group PG^(2a) thatis selectively cleavable towards PG², and the hydroxy function may beconverted to a group —OPG¹ or to a group —O—R⁹. The products obtainedupon removal of the group PG² are intermediates (17 g) and (17i) whichcorrespond to intermediates (13a) or (16a) specified above.

Intermediates with specific stereochemistry may be prepared by resolvingthe intermediates in the above reaction sequence. For example, (17b) maybe resolved following art-known procedures, e.g. by salt form actionwith an optically active base or by chiral chromatography, and theresulting stereoisomers may be processed further as described above. TheOH and COOH groups in (17d) are in cis position. Trans analogs can beprepared by inverting the stereochemistry at the carbon bearing the OHfunction by using specific reagents in the reactions introducing OPG¹ orO—R⁹ that invert the stereochemistry, such as, e.g. by applying aMitsunobu reaction.

In one embodiment, the intermediates (17d) are coupled to P1 blocks(12b) or (12c), which coupling reactions correspond to the coupling of(13a) or (16a) with the same P1 blocks, using the same conditions.Subsequent introduction of a —O—R⁹ substituent as described abovefollowed by removal of the acid protection group PG² yieldsintermediates (8a-1), which are a subclass of the intermediates (7a), orpart of the intermediates (16a). The reaction products of the PG²removal can be further coupled to a P3 building block. In one embodimentPG² in (17d) is t.butyl which can be removed under acidic conditions,e.g. with trifluoroacetic acid.

An unsaturated P2 building block, i.e. a cyclopentene ring, may beprepared as illustrated in the scheme below.

A bromination-elimination reaction of3,4-bis(methoxycarbonyl)cyclopentanone (17a) as described by Dolby etal. in J. Org. Chem. 36 (1971) 1277-1285 followed by reduction of theketo functionality with a reducting agent like sodium borohydrideprovides the cyclopentenol (19a). Selective ester hydrolysis using forexample lithium hydroxide in a solvent like a mixture of dioxane andwater, provides the hydroxy substituted monoester cyclopentenol (19b).

An unsaturated P2 building block wherein R² can also be other thanhydrogen, may be prepared as shown in the scheme below.

Oxidation of commercially available 3-methyl-3-buten-1-ol (20a), inparticular by an oxidizing agent like pyridinium chlorochromate, yields(20b), which is converted to the corresponding methyl ester, e.g. bytreatment with acetyl chloride in methanol, followed by a brominationreaction with bromine yielding the α-bromo ester (20c). The latter canthen be condensed with the alkenyl ester (20e), obtained from (20d) byan ester forming reaction. The ester in (20e) preferably is a t.butylester which can be prepared from the corresponding commerciallyavailable acid (20d), e.g. by treatment with di-tert-butyl dicarbonatein the presence of a base like dimethylaminopyridine.

Intermediate (20e) is treated with a base such as lithium diisopropylamide in a solvent like tetrahydrofuran, and reacted with (20c) to givethe alkenyl diester (20f). Cyclisation of (20f) by an olefin metathesisreaction, performed as described above, provides cyclopentene derivative(20 g). Stereoselective epoxidation of (20 g) can be carried out usingthe Jacobsen asymmetric epoxidation method to obtain epoxide (20 h).Finally, an epoxide opening reaction under basic conditions, e.g. byaddition of a base, in particular DBN(1,5-diazabicyclo-[4.3.0]non-5-ene), yields the alcohol (20i).Optionally, the double bond in intermediate (20i) can be reduced, forexample by catalytic hydrogenation using a catalyst like palladium oncarbon, yielding the corresponding cyclopentane compound. The t.butylester may be removed to the corresponding acid, which subsequently iscoupled to a P1 building block.

The —R⁹ group can be introduced on the pyrrolidine, cyclopentane orcyclopentene rings at any convenient stage of the synthesis of thecompounds according to the present invention. One approach is to firstintroduce the —R⁹ group to the said rings and subsequently add the otherdesired building blocks, i.e. P1 (optionally with the P1′ tail) and P3,followed by the macrocycle formation. Another approach is to couple thebuilding blocks P2, bearing no —O—R⁹ substituent, with each P1 and P3,and to add the —R⁹ group either before or after the macrocycleformation. In the latter procedure, the P2 moieties have a hydroxygroup, which may be protected by a hydroxy protecting group PG¹.

R⁹ groups can be introduced on building blocks P2 by reacting hydroxysubstituted intermediates (21a) or (21b) with intermediates (4b) similaras described above for the synthesis of (I) starting from (4a). Thesereactions are represented in the schemes below, wherein L² is asspecified above and L⁵ and L^(5a) independently from one another,represent hydroxy, a carboxyl protecting group —OPG² or —OPG^(2a), or L⁵may also represent a P1 group such as a group (d) or (e) as specifiedabove, or L^(5a) may also represent a P3 group such as a group (b) asspecified above The groups PG² and PG^(2a) are as specified above. Wherethe groups L⁵ and L^(5a) are PG² or PG^(2a), they are chosen such thateach group is selectively cleavable towards the other. For example, oneof L⁵ and L^(5a) may be a methyl or ethyl group and the other a benzylor t.butyl group.

In one embodiment in (21a), L² is PG and L⁵ is —OPG², or in (21d),L^(5a) is —OPG² and L⁵ is —OPG² and the PG² groups are removed asdescribed above.

Alternatively, when handling hydroxy substituted cyclopentane analogues,the quinoline substituent can be introduced via a similar Mitsunobureaction by reacting the hydroxy group of compound (2a′) with thedesired alcohol (3b) in the presence of triphenylphosphine and anactivating agent like diethyl azodicarboxylate (DEAD), diisopropylazodicarboxylate (DIAD) or the like.

In another embodiment the group L² is BOC, L⁵ is hydroxy and thestarting material (21a) is commercially available BOC-hydroxyproline, orany other stereoisomeric form thereof, e.g. BOC-L-hydroxyproline, inparticular the trans isomer of the latter. Where L⁵ in (21b) is acarboxyl-protecting group, it may be removed following proceduresdescribed above to (21c). In still another embodiment PG in (21b-1) isBoc and PG² is a lower alkyl ester, in particular a methyl or ethylester. Hydrolysis of the latter ester to the acid can be done bystandard procedures, e.g. acid hydrolysis with hydrochloric acid inmethanol or with an alkali metal hydroxide such as NaOH, in particularwith LiOH. In another embodiment, hydroxy substituted cyclopentane orcyclopentene analogs (21d) are converted to (21e), which, where L⁵ andL^(5a) are —OPG² or —OPG^(2a), may be converted to the correspondingacids (21f) by removal of the group PG². Removal of PG^(2a) in (21e-1)leads to similar intermediates.

The intermediates Y—R⁹ (4b) can be prepared following art-known methodsusing known starting materials. A number of synthesis pathways for suchintermediates will be described hereafter in somewhat more detail. Forexample the preparation of the above mentioned intermediate quinolinesis shown below in the following scheme.

Friedel-Craft acylation of a suitable substituted aniline (22a),available either commercially or via art-known procedures, using anacylating agent such as acetyl chloride or the like in the presence ofone or more Lewis acid such as boron trichloride and aluminumtrichloride in a solvent like dichloromethane provides (22b). Couplingof (22b) with a carboxylic acid (22c), preferably under basicconditions, such as in pyridine, in the presence of an activating agentfor the carboxylate group, for instance POCl₃, followed by ring closureand dehydration under basic conditions like potassium tert-butoxide intert-butanol yields quinoline derivative (22e). The latter can beconverted to (22f) wherein LG is a leaving group, e.g. by reaction of(22e) with a halogenating agent, for example phosphoryl chloride or thelike, or with an arylsulfonyl chloride, e.g. with tosyl chloride.Quinoline derivative (22e) can be coupled in a Mitsunobu reaction to analcohol as described above, or quinoline (22f) can be reacted with (1a)in an O-arylation reaction as described above.

A variety of carboxylic acids with the general structure (22c) can beused in the above synthesis. These acids are available eithercommercially or can be prepared via art-known procedures. An example ofthe preparation of 2-(substituted)aminocarboxy-aminothiazole derivatives(23a-1), following the procedure described by Berdikhina et al. in Chem.Heterocycl. Compd. (Engl. Transl.) (1991), 427-433, is shown in thefollowing reaction scheme which illustrates the preparation of2-carboxy-4-isopropyl-thiazole (22c-1):

Ethyl thiooxamate (23a) is reacted with the β-bromoketone (23b) to formthe thiazolyl carboxylic acid ester (23c), which is hydrolyzed to thecorresponding acid (25c-1). The ethyl ester in these intermediates maybe replaced by other carboxyl protecting groups PG², as defined above.In the above scheme R^(4a) is as defined above and in particular isC₁₋₄alkyl, more in particular i.propyl.

The bromoketone (23b) may be prepared from 3-methyl-butan-2-one (MIK)with a sililating agent (such as TMSCl) in the presence of a suitablebase (in particular LiHMDS) and bromine

The synthesis of further carboxylic acids (22c), in particular ofsubstituted amino thiazole carboxylic acids (25a-2) is illustratedherebelow:

Thiourea (24c) with various substituents R^(4a), which in particular areC₁₋₆alkyl, can be formed by reaction of the appropriate amine (24a) withtert-butylisothiocyanate in the presence of a base likediisopropylethylamine in a solvent like dichloromethane followed byremoval of the tert-butyl group under acidic conditions. Subsequentcondensation of thiourea derivative (24c) with 3-bromopyruvic acidprovides the thiazole carboxylic acid (22c-2).

Synthesis of P1 Building Blocks

The cyclopropane amino acid used in the preparation of the P1 fragmentis commercially available or can be prepared using art-known procedures.

In particular the amino-vinyl-cyclopropyl ethyl ester (12b) may beobtained according to the procedure described in WO 00/09543 or asillustrated in the following scheme, wherein PG² is a carboxylprotecting group as specified above:

Treatment of commercially available or easily obtainable imine (25a)with 1,4-dihalo-butene in presence of a base produces (25b), which afterhydrolysis yields cyclopropyl amino acid (12b), having the allylsubstituent syn to the carboxyl group. Resolution of the enantiomericmixture (12b) results in (12b-1). The resolution is performed usingart-known procedures such as enzymatic separation; crystallization witha chiral acid; or chemical derivatization; or by chiral columnchromatography. Intermediates (12b) or (12b-1) may be coupled to theappropriate P2 derivatives as described above.

P1 building blocks for the preparation of compounds according to generalformula (I) wherein R¹ is —OR⁷ or —NH—SO₂R⁸ can be prepared by reactingamino acids (23a) with the appropriate alcohol or amine respectivelyunder standard conditions for ester or amide formation. Cyclopropylamino acids (23a) are prepared by introducing a N-protecting group PG,and removal of PG² and the amino acids (a) are converted to the amides(12c-1) or esters (12c-2), which are subgroups of the intermediates(12c), as outlined in the following reaction scheme, wherein PG is asspecified above.

The reaction of (26a) with amine (2b) is an amide forming procedure. Thesimilar reaction with (2c) is an ester forming reaction. Both can beperformed following the procedures described above. This reaction yieldsintermediates (26b) or (26c) from which the amino protecting group isremoved by standard methods such as those described above. This in turnresults in the desired intermediate (12c-1). Starting materials (26a)may be prepared from the above-mentioned intermediates (12b) by firstintroducing a N-protecting group PG and subsequent removal of the groupPG².

In one embodiment the reaction of (26a) with (2b) is done by treatmentof the amino acid with a coupling agent, for exampleN,N′-carbonyl-diimidazole (CDI) or the like, in a solvent like THFfollowed by reaction with (2b) in the presence of a base such as1,8-diazabicyclo[5.4.0]undec-7-ene (DBU). Alternatively the amino acidcan be treated with (2b) in the presence of a base likediisopropylethylamine followed by treatment with a coupling agent suchas benzotriazole-1-yl-oxy-tris-pyrrolidino-phosphoniumhexafluorophosphate (commercially available as PyBOP®) to effect theintroduction of the sulfonamide group.

Intermediates (12c-1) or (12c-2) in turn may be coupled to theappropriate proline, cyclopentane or cyclopentene derivatives asdescribed above.

Synthesis of the P3 Building Blocks

The P3 building blocks are available commercially or can be preparedaccording to methodologies known to the skilled in the art. One of thesemethodologies is shown in the scheme below and uses monoacylated amines,such as trifluoroacetamide or a Boc-protected amine

In the above scheme, R together with the CO group forms a N-protectinggroup, in particular R is t-butoxy, trifluoromethyl; R³ and n are asdefined above and LG is a leaving group, in particular halogen, e.g.chloro or bromo.

The monoacylated amines (27a) are treated with a strong base such assodium hydride and are subsequently reacted with a reagentLG-C₅₋₈alkenyl (27b), in particular haloC₅₋₈alkenyl, to form thecorresponding protected amines (27c). Deprotection of (27c) affords(5b), which are building blocks P3. Deprotection will depend on thefunctional group R, thus if R is t-butoxy, deprotection of thecorresponding Boc-protected amine can be accomplished with an acidictreatment, e.g. trifluoroacetic acid. Alternatively, when R is forinstance trifluoromethyl, removal of the R group is accomplished with abase, e.g. sodium hydroxide.

The following scheme illustrates yet another method for preparing a P3building block, namely a Gabriel synthesis of primary C₅₋₈alkenylamines,which can be carried out by the treatment of a phthalimide (28a) with abase, such as NaOH or KOH, and with (27b), which is as specified above,followed by hydrolysis of the intermediate N-alkenyl imide to generate aprimary C₅₋₈alkenylamine (5b-1).

In the above scheme, n is as defined above.

Compounds of formula (I) may be converted into each other followingart-known functional group transformation reactions. For example, aminogroups may be N-alkylated, nitro groups reduced to amino groups, a haloatom may be exchanged for another halo.

The compounds of formula (I) may be converted to the correspondingN-oxide forms following art-known procedures for converting a trivalentnitrogen into its N-oxide form. Said N-oxidation reaction may generallybe carried out by reacting the starting material of formula (I) with anappropriate organic or inorganic peroxide. Appropriate inorganicperoxides comprise, for example, hydrogen peroxide, alkali metal orearth alkaline metal peroxides, e.g. sodium peroxide, potassiumperoxide; appropriate organic peroxides may comprise peroxy acids suchas, for example, benzenecarbo-peroxoic acid or halo substitutedbenzenecarboperoxoic acid, e.g. 3-chlorobenzene-carboperoxoic acid,peroxoalkanoic acids, e.g. peroxoacetic acid, alkylhydroperoxides, e.g.tert-butyl hydro-peroxide. Suitable solvents are, for example, water,lower alcohols, e.g. ethanol and the like, hydrocarbons, e.g. toluene,ketones, e.g. 2-butanone, halogenated hydrocarbons, e.g.dichloromethane, and mixtures of such solvents.

Pure stereochemically isomeric forms of the compounds of formula (I) maybe obtained by the application of art-known procedures. Diastereomersmay be separated by physical methods such as selective crystallizationand chromatographic techniques, e.g., counter-current distribution,liquid chromatography and the like.

The compounds of formula (I) may be obtained as racemic mixtures ofenantiomers which can be separated from one another following art-knownresolution procedures. The racemic compounds of formula (I), which aresufficiently basic or acidic may be converted into the correspondingdiastereomeric salt forms by reaction with a suitable chiral acid,respectively chiral base. Said diastereomeric salt forms aresubsequently separated, for example, by selective or fractionalcrystallization and the enantiomers are liberated therefrom by alkali oracid. An alternative manner of separating the enantiomeric forms of thecompounds of formula (I) involves liquid chromatography, in particularliquid chromatography using a chiral stationary phase. Said purestereochemically isomeric forms may also be derived from thecorresponding pure stereochemically isomeric forms of the appropriatestarting materials, provided that the reaction occursstereospecifically. Preferably if a specific stereoisomer is desired,said compound may be synthesized by stereospecific methods ofpreparation. These methods may advantageously employ enantiomericallypure starting materials.

In a further aspect, the present invention concerns a pharmaceuticalcomposition comprising a therapeutically effective amount of a compoundof formula (I) as specified herein, or a compound of any of thesubgroups of compounds of formula (I) as specified herein, and apharmaceutically acceptable carrier. A therapeutically effective amountin this context is an amount sufficient to prophylactically act against,to stabilize or to reduce viral infection, and in particular HCV viralinfection, in infected subjects or subjects being at risk of beinginfected. In still a further aspect, this invention relates to a processof preparing a pharmaceutical composition as specified herein, whichcomprises intimately mixing a pharmaceutically acceptable carrier with atherapeutically effective amount of a compound of formula (I), asspecified herein, or of a compound of any of the subgroups of compoundsof formula (I) as specified herein.

Therefore, the compounds of the present invention or any subgroupthereof may be formulated into various pharmaceutical forms foradministration purposes. As appropriate compositions there may be citedall compositions usually employed for systemically administering drugs.To prepare the pharmaceutical compositions of this invention, aneffective amount of the particular compound, optionally in addition saltform or metal complex, as the active ingredient is combined in intimateadmixture with a pharmaceutically acceptable carrier, which carrier maytake a wide variety of forms depending on the form of preparationdesired for administration. These pharmaceutical compositions aredesirable in unitary dosage form suitable, particularly, foradministration orally, rectally, percutaneously, or by parenteralinjection. For example, in preparing the compositions in oral dosageform, any of the usual pharmaceutical media may be employed such as, forexample, water, glycols, oils, alcohols and the like in the case of oralliquid preparations such as suspensions, syrups, elixirs, emulsions andsolutions; or solid carriers such as starches, sugars, kaolin,lubricants, binders, disintegrating agents and the like in the case ofpowders, pills, capsules, and tablets. Because of their ease inadministration, tablets and capsules represent the most advantageousoral dosage unit forms, in which case solid pharmaceutical carriers areobviously employed. For parenteral compositions, the carrier willusually comprise sterile water, at least in large part, though otheringredients, for example, to aid solubility, may be included. Injectablesolutions, for example, may be prepared in which the carrier comprisessaline solution, glucose solution or a mixture of saline and glucosesolution. Injectable suspensions may also be prepared in which caseappropriate liquid carriers, suspending agents and the like may beemployed. Also included are solid form preparations which are intendedto be converted, shortly before use, to liquid form preparations. In thecompositions suitable for percutaneous administration, the carrieroptionally comprises a penetration enhancing agent and/or a suitablewetting agent, optionally combined with suitable additives of any naturein minor proportions, which additives do not introduce a significantdeleterious effect on the skin.

The compounds of the present invention may also be administered via oralinhalation or insufflation by means of methods and formulations employedin the art for administration via this way. Thus, in general thecompounds of the present invention may be administered to the lungs inthe form of a solution, a suspension or a dry powder, a solution beingpreferred. Any system developed for the delivery of solutions,suspensions or dry powders via oral inhalation or insufflation aresuitable for the administration of the present compounds.

Thus, the present invention also provides a pharmaceutical compositionadapted for administration by inhalation or insufflation through themouth comprising a compound of formula (I) and a pharmaceuticallyacceptable carrier. Preferably, the compounds of the present inventionare administered via inhalation of a solution in nebulized oraerosolized doses.

It is especially advantageous to formulate the aforementionedpharmaceutical compositions in unit dosage form for ease ofadministration and uniformity of dosage. Unit dosage form as used hereinrefers to physically discrete units suitable as unitary dosages, eachunit containing a predetermined quantity of active ingredient calculatedto produce the desired therapeutic effect in association with therequired pharmaceutical carrier. Examples of such unit dosage forms aretablets (including scored or coated tablets), capsules, pills,suppositories, powder packets, wafers, injectable solutions orsuspensions and the like, and segregated multiples thereof.

The compounds of formula (I) show antiviral properties. Viral infectionsand their associated diseases treatable using the compounds and methodsof the present invention include those infections brought on by HCV andother pathogenic flaviviruses such as Yellow fever, Dengue fever (types1-4), St. Louis encephalitis, Japanese encephalitis, Murray valleyencephalitis, West Nile virus and Kunjin virus. The diseases associatedwith HCV include progressive liver fibrosis, inflammation and necrosisleading to cirrhosis, end-stage liver disease, and HCC; and for theother pathogenic flaviviruses the diseases include yellow fever, denguefever, hemorrhagic fever and encephalitis. A number of the compounds ofthis invention moreover are active against mutated strains of HCV.Additionally, many of the compounds of this invention show a favorablepharmacokinetic profile and have attractive properties in terms ofbioavailabilty, including an acceptable half-life, AUC (area under thecurve) and peak values and lacking unfavorable phenomena such asinsufficient quick onset and tissue retention.

The in vitro antiviral activity against HCV of the compounds of formula(I) was tested in a cellular HCV replicon system based on Lohmann et al.(1999) Science 285:110-113, with the further modifications described byKrieger et al. (2001) Journal of Virology 75: 4614-4624, which isfurther exemplified in the examples section. This model, while not acomplete infection model for HCV, is widely accepted as the most robustand efficient model of autonomous HCV RNA replication currentlyavailable. Compounds exhibiting anti-HCV activity in this cellular modelare considered as candidates for further development in the treatment ofHCV infections in mammals. It will be appreciated that it is importantto distinguish between compounds that specifically interfere with HCVfunctions from those that exert cytotoxic or cytostatic effects in theHCV replicon model, and as a consequence cause a decrease in HCV RNA orlinked reporter enzyme concentration. Assays are known in the field forthe evaluation of cellular cytotoxicity based for example on theactivity of mitochondrial enzymes using fluorogenic redox dyes such asresazurin. Furthermore, cellular counter screens exist for theevaluation of non-selective inhibition of linked reporter gene activity,such as firefly luciferase. Appropriate cell types can be equipped bystable transfection with a luciferase reporter gene whose expression isdependent on a constitutively active gene promoter, and such cells canbe used as a counter-screen to eliminate non-selective inhibitors.

Due to their antiviral properties, particularly their anti-HCVproperties, the compounds of formula (I) or any subgroup thereof, theirprodrugs, N-oxides, addition salts, quaternary amines, metal complexesand stereochemically isomeric forms, are useful in the treatment ofindividuals experiencing a viral infection, particularly a HCVinfection, and for the prophylaxis of these infections. In general, thecompounds of the present invention may be useful in the treatment ofwarm-blooded animals infected with viruses, in particular flavivirusessuch as HCV.

The compounds of the present invention or any subgroup thereof maytherefore be used as medicines. Said use as a medicine or method oftreatment comprises the systemic administration to viral infectedsubjects or to subjects susceptible to viral infections of an amounteffective to combat the conditions associated with the viral infection,in particular the HCV infection.

The present invention also relates to the use of the present compoundsor any subgroup thereof in the manufacture of a medicament for thetreatment or the prevention of viral infections, particularly HCVinfection.

The present invention furthermore relates to a method of treating awarm-blooded animal infected by a virus, or being at risk of infectionby a virus, in particular by HCV, said method comprising theadministration of an anti-virally effective amount of a compound offormula (I), as specified herein, or of a compound of any of thesubgroups of compounds of formula (I), as specified herein.

Also, the combination of previously known anti-HCV compound, such as,for instance, interferon-α (IFN-α), pegylated interferon-α and/orribavirin, and a compound of formula (I) can be used as a medicine in acombination therapy. The term “combination therapy” relates to a productcontaining mandatory (a) a compound of formula (I), and (b) optionallyanother anti-HCV compound, as a combined preparation for simultaneous,separate or sequential use in treatment of HCV infections, inparticular, in the treatment of infections with HCV.

Anti-HCV compounds encompass agents selected from an HCV polymeraseinhibitor, an HCV protease inhibitor, an inhibitor of another target inthe HCV life cycle, and immunomodulatory agent, an antiviral agent, andcombinations thereof.

HCV polymerase inhibitors include, but are not limited to, NM283(valopicitabine), R803, JTK-109, JTK-003, HCV-371, HCV-086, HCV-796 andR-1479.

Inhibitors of HCV proteases (NS2-NS3 inhibitors and NS3-NS4A inhibitors)include, but are not limited to, the compounds of WO02/18369 (see, e.g.,page 273, lines 9-22 and page 274, line 4 to page 276, line 11);BILN-2061, VX-950, GS-9132 (ACH-806), SCH-503034, and SCH-6. Furtheragents that can be used are those disclosed in WO-98/17679, WO-00/056331(Vertex); WO 98/22496 (Roche); WO 99/07734, (Boehringer Ingelheim), WO2005/073216, WO2005073195 (Medivir) and structurally similar agents.

Inhibitors of other targets in the HCV life cycle, including NS3helicase; metallo-protease inhibitors; antisense oligonucleotideinhibitors, such as ISIS-14803, AVI-4065 and the like; siRNA's such asSIRPLEX-140-N and the like; vector-encoded short hairpin RNA (shRNA);DNAzymes; HCV specific ribozymes such as heptazyme, RPI.13919 and thelike; entry inhibitors such as HepeX-C, HuMax-HepC and the like; alphaglucosidase inhibitors such as celgosivir, UT-231B and the like;KPE-02003002; and BIVN 401.

Immunomodulatory agents include, but are not limited to; natural andrecombinant interferon isoform compounds, including α-interferon,β-interferon, γ-interferon, ω-interferon and the like, such as IntronA®, Roferon-A®, Canferon-A300®, Advaferon®, Infergen®, Humoferon®,Sumiferon MP®, Alfaferone®, IFN-beta®, Feron® and the like; polyethyleneglycol derivatized (pegylated) interferon compounds, such as PEGinterferon-α-2a (Pegasys®), PEG interferon-α-2b (PEG-Intron®), pegylatedIFN-α-con1 and the like; long acting formulations and derivatizations ofinterferon compounds such as the albumin-fused interferon albuferon αand the like; compounds that stimulate the synthesis of interferon incells, such as resiquimod and the like; interleukins; compounds thatenhance the development of type 1 helper T cell response, such as SCV-07and the like; TOLL-like receptor agonists such as CpG-10101 (actilon),isatoribine and the like; thymosin α-1; ANA-245; ANA-246; histaminedihydrochloride; propagermanium; tetrachlorodecaoxide; ampligen;IMP-321; KRN-7000; antibodies, such as civacir, XTL-6865 and the like;and prophylactic and therapeutic vaccines such as InnoVac C, HCVE1E2/MF59 and the like.

Other antiviral agents include, but are not limited to, ribavirin,amantadine, viramidine, nitazoxanide; telbivudine; NOV-205; taribavirin;inhibitors of internal ribosome entry; broad-spectrum viral inhibitors,such as IMPDH inhibitors (e.g., compounds of U.S. Pat. No. 5,807,876,U.S. Pat. No. 6,498,178, U.S. Pat. No. 6,344,465, U.S. Pat. No.6,054,472, WO97/40028, WO98/40381, WO00/56331, and mycophenolic acid andderivatives thereof, and including, but not limited to VX-950,merimepodib (VX-497), VX-148, and/or VX-944); or combinations of any ofthe above.

Thus, to combat or treat HCV infections, the compounds of formula (I)may be co-administered in combination with for instance, interferon-α(IFN-α), pegylated interferon-α and/or ribavirin, as well astherapeutics based on antibodies targeted against HCV epitopes, smallinterfering RNA (Si RNA), ribozymes, DNAzymes, antisense RNA, smallmolecule antagonists of for instance NS3 protease, NS3 helicase and NS5Bpolymerase.

Accordingly, the present invention relates to the use of a compound offormula (I) or any subgroup thereof as defined above for the manufactureof a medicament useful for inhibiting HCV activity in a mammal infectedwith HCV viruses, wherein said medicament is used in a combinationtherapy, said combination therapy preferably comprising a compound offormula (I) and another HCV inhibitory compound, e.g. (pegylated) IFN-αand/or ribavirin.

In still another aspect there are provided combinations of a compound offormula (I) as specified herein and an anti-HIV compound. The latterpreferably are those HIV inhibitors that have a positive effect on drugmetabolism and/or pharmacokinetics that improve bioavailabilty. Anexample of such an HIV inhibitor is ritonavir.

As such, the present invention further provides a combination comprising(a) an HCV NS3/4a protease inhibitor of formula (I) or apharmaceutically acceptable salt thereof; and (b) ritonavir or apharmaceutically acceptable salt thereof.

The compound ritonavir, and pharmaceutically acceptable salts thereof,and methods for its preparation are described in WO94/14436. Forpreferred dosage forms of ritonavir, see U.S. Pat. No. 6,037,157, andthe documents cited therein: U.S. Pat. No. 5,484,801, U.S. Ser. No.08/402,690, and WO95/07696 and WO95/09614. Ritonavir has the followingformula:

In a further embodiment, the combination comprising (a) an HCV NS3/4aprotease inhibitor of formula (I) or a pharmaceutically acceptable saltthereof; and (b) ritonavir or a pharmaceutically acceptable saltthereof; further comprises an additional anti-HCV compound selected fromthe compounds as described herein.

In one embodiment of the present invention there is provided a processfor preparing a combination as described herein, comprising the step ofcombining an HCV NS3/4a protease inhibitor of formula (I) or apharmaceutically acceptable salt thereof, and ritonavir or apharmaceutically acceptable salt thereof. An alternative embodiment ofthis invention provides a process wherein the combination comprises oneor more additional agent as described herein.

The combinations of the present invention may be used as medicaments.Said use as a medicine or method of treatment comprises the systemicadministration to HCV-infected subjects of an amount effective to combatthe conditions associated with HCV and other pathogenic flavi- andpestiviruses. Consequently, the combinations of the present inventioncan be used in the manufacture of a medicament useful for treating,preventing or combating infection or disease associated with HCVinfection in a mammal, in particular for treating conditions associatedwith HCV and other pathogenic flavi- and pestiviruses.

In one embodiment of the present invention there is provided apharmaceutical composition comprising a combination according to any oneof the embodiments described herein and a pharmaceutically acceptableexcipient. In particular, the present invention provides apharmaceutical composition comprising (a) a therapeutically effectiveamount of an HCV NS3/4a protease inhibitor of the formula (I) or apharmaceutically acceptable salt thereof, (b) a therapeuticallyeffective amount of ritonavir or a pharmaceutically acceptable saltthereof, and (c) a pharmaceutically acceptable excipient. Optionally,the pharmaceutical composition further comprises an additional agentselected from an HCV polymerase inhibitor, an HCV protease inhibitor, aninhibitor of another target in the HCV life cycle, and immunomodulatoryagent, an antiviral agent, and combinations thereof.

The compositions may be formulated into suitable pharmaceutical dosageforms such as the dosage forms described above. Each of the activeingredients may be formulated separately and the formulations may beco-administered or one formulation containing both and if desiredfurther active ingredients may be provided.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients, as well as any productwhich results, directly or indirectly, from the combination of thespecified ingredients.

In one embodiment the combinations provided herein may also beformulated as a combined preparation for simultaneous, separate orsequential use in HIV therapy. In such a case, the compound of generalformula (I) or any subgroup thereof, is formulated in a pharmaceuticalcomposition containing other pharmaceutically acceptable excipients, andritonavir is formulated separately in a pharmaceutical compositioncontaining other pharmaceutically acceptable excipients. Conveniently,these two separate pharmaceutical compositions can be part of a kit forsimultaneous, separate or sequential use.

Thus, the individual components of the combination of the presentinvention can be administered separately at different times during thecourse of therapy or concurrently in divided or single combinationforms. The present invention is therefore to be understood as embracingall such regimes of simultaneous or alternating treatment and the term“administering” is to be interpreted accordingly. In a preferredembodiment, the separate dosage forms are administered aboutsimultaneously.

In one embodiment, the combination of the present invention contains anamount of ritonavir, or a pharmaceutically acceptable salt thereof,which is sufficient to clinically improve the bioavailability of the HCVNS3/4a protease inhibitor of formula (I) relative to the bioavailabilitywhen said HCV NS3/4a protease inhibitor of formula (I) is administeredalone.

In another embodiment, the combination of the present invention containsan amount of ritonavir, or a pharmaceutically acceptable salt thereof,which is sufficient to increase at least one of the pharmacokineticvariables of the HCV NS3/4a protease inhibitor of formula (I) selectedfrom t_(1/2), C_(min), C_(max), C_(ss), AUC at 12 hours, or AUC at 24hours, relative to said at least one pharmacokinetic variable when theHCV NS3/4a protease inhibitor of formula (I) is administered alone.

A further embodiment relates to a method for improving thebioavailability of a HCV NS3/4a protease inhibitor comprisingadministering to an individual in need of such improvement a combinationas defined herein, comprising a therapeutically effective amount of eachcomponent of said combination.

In a further embodiment, the invention relates to the use of ritonaviror a pharmaceutically acceptable salt thereof, as an improver of atleast one of the pharmacokinetic variables of a HCV NS3/4a proteaseinhibitor of formula (I) selected from t_(1/2), C_(min), C_(max),C_(ss), AUC at 12 hours, or AUC at 24 hours; with the proviso that saiduse is not practised in the human or animal body.

The term “individual” as used herein refers to an animal, preferably amammal, most preferably a human, who has been the object of treatment,observation or experiment.

Bioavailability is defined as the fraction of administered dose reachingsystemic circulation. t_(1/2) represents the half life or time taken forthe plasma concentration to fall to half its original value. C_(ss) isthe steady state concentration, i.e. the concentration at which the rateof input of drug equals the rate of elimination. C_(min) is defined asthe lowest (minimum) concentration measured during the dosing interval.C_(max), represents the highest (maximum) concentration measured duringthe dosing interval. AUC is defined as the area under the plasmaconcentration-time curve for a defined period of time.

The combinations of this invention can be administered to humans indosage ranges specific for each component comprised in saidcombinations. The components comprised in said combinations can beadministered together or separately. The NS3/4a protease inhibitors offormula (I) or any subgroup thereof, and ritonavir or a pharmaceuticallyacceptable salt or ester thereof, may have dosage levels of the order of0.02 to 5.0 grams-per-day.

When the HCV NS3/4a protease inhibitor of formula (I) and ritonavir areadministered in combination, the weight ratio of the HCV NS3/4a proteaseinhibitor of formula (I) to ritonavir is suitably in the range of fromabout 40:1 to about 1:15, or from about 30:1 to about 1:15, or fromabout 15:1 to about 1:15, typically from about 10:1 to about 1:10, andmore typically from about 8:1 to about 1:8. Also useful are weightratios of the HCV NS3/4a protease inhibitors of formula (I) to ritonavirranging from about 6:1 to about 1:6, or from about 4:1 to about 1:4, orfrom about 3:1 to about 1:3, or from about 2:1 to about 1:2, or fromabout 1.5:1 to about 1:1.5. In one aspect, the amount by weight of theHCV NS3/4a protease inhibitors of formula (I) is equal to or greaterthan that of ritonavir, wherein the weight ratio of the HCV NS3/4aprotease inhibitor of formula (I) to ritonavir is suitably in the rangeof from about 1:1 to about 15:1, typically from about 1:1 to about 10:1,and more typically from about 1:1 to about 8:1. Also useful are weightratios of the HCV NS3/4a protease inhibitor of formula (I) to ritonavirranging from about 1:1 to about 6:1, or from about 1:1 to about 5:1, orfrom about 1:1 to about 4:1, or from about 3:2 to about 3:1, or fromabout 1:1 to about 2:1 or from about 1:1 to about 1.5:1.

The term “therapeutically effective amount” as used herein means thatamount of active compound or component or pharmaceutical agent thatelicits the biological or medicinal response in a tissue, system, animalor human that is being sought, in the light of the present invention, bya researcher, veterinarian, medical doctor or other clinician, whichincludes alleviation of the symptoms of the disease being treated. Sincethe instant invention refers to combinations comprising two or moreagents, the “therapeutically effective amount” is that amount of theagents taken together so that the combined effect elicits the desiredbiological or medicinal response. For example, the therapeuticallyeffective amount of a composition comprising (a) the compound of formula(I) and (b) ritonavir, would be the amount of the compound of formula(I) and the amount of ritonavir that when taken together have a combinedeffect that is therapeutically effective.

In general it is contemplated that an antiviral effective daily amountwould be from 0.01 mg/kg to 500 mg/kg body weight, more preferably from0.1 mg/kg to 50 mg/kg body weight. It may be appropriate to administerthe required dose as two, three, four or more sub-doses at appropriateintervals throughout the day. Said sub-doses may be formulated as unitdosage forms, for example, containing 1 to 1000 mg, and in particular 5to 200 mg of active ingredient per unit dosage form.

The exact dosage and frequency of administration depends on theparticular compound of formula (I) used, the particular condition beingtreated, the severity of the condition being treated, the age, weight,sex, extent of disorder and general physical condition of the particularpatient as well as other medication the individual may be taking, as iswell known to those skilled in the art. Furthermore, it is evident thatsaid effective daily amount may be lowered or increased depending on theresponse of the treated subject and/or depending on the evaluation ofthe physician prescribing the compounds of the instant invention. Theeffective daily amount ranges mentioned hereinabove are therefore onlyguidelines.

According to one embodiment, the HCV NS3/4a protease inhibitor offormula (I) and ritonavir may be co-administered once or twice a day,preferably orally, wherein the amount of the compounds of formula (I)per dose is from about 1 to about 2500 mg, and the amount of ritonavirper dose is from Ito about 2500 mg. In another embodiment, the amountsper dose for once or twice daily co-administration are from about 50 toabout 1500 mg of the compound of formula (I) and from about 50 to about1500 mg of ritonavir. In still another embodiment, the amounts per dosefor once or twice daily co-administration are from about 100 to about1000 mg of the compound of formula (I) and from about 100 to about 800mg of ritonavir. In yet another embodiment, the amounts per dose foronce or twice daily co-administration are from about 150 to about 800 mgof the compound of formula (I) and from about 100 to about 600 mg ofritonavir. In yet another embodiment, the amounts per dose for once ortwice daily co-administration are from about 200 to about 600 mg of thecompound of formula (I) and from about 100 to about 400 mg of ritonavir.In yet another embodiment, the amounts per dose for once or twice dailyco-administration are from about 200 to about 600 mg of the compound offormula (I) and from about 20 to about 300 mg of ritonavir. In yetanother embodiment, the amounts per dose for once or twice dailyco-administration are from about 100 to about 400 mg of the compound offormula (I) and from about 40 to about 100 mg of ritonavir.

Exemplary combinations of the compound of formula (I) (mg)/ritonavir(mg) for once or twice daily dosage include 50/100, 100/100, 150/100,200/100, 250/100, 300/100, 350/100, 400/100, 450/100, 50/133, 100/133,150/133, 200/133, 250/133, 300/133, 50/150, 100/150, 150/150, 200/150,250/150, 50/200, 100/200, 150/200, 200/200, 250/200, 300/200, 50/300,80/300, 150/300, 200/300, 250/300, 300/300, 200/600, 400/600, 600/600,800/600, 1000/600, 200/666, 400/666, 600/666, 800/666, 1000/666,1200/666, 200/800, 400/800, 600/800, 800/800, 1000/800, 1200/800,200/1200, 400/1200, 600/1200, 800/1200, 1000/1200, and 1200/1200. Otherexemplary combinations of the compound of formula (I) (mg)/ritonavir(mg) for once or twice daily dosage include 1200/400, 800/400, 600/400,400/200, 600/200, 600/100, 500/100, 400/50, 300/50, and 200/50.

In one embodiment of the present invention there is provided an articleof manufacture comprising a composition effective to treat an HCVinfection or to inhibit the NS3 protease of HCV; and packaging materialcomprising a label which indicates that the composition can be used totreat infection by the hepatitis C virus; wherein the compositioncomprises a compound of the formula (I) or any subgroup thereof, or thecombination as described herein.

Another embodiment of the present invention concerns a kit or containercomprising a compound of the formula (I) or any subgroup thereof, or acombination according to the invention combining an HCV NS3/4a proteaseinhibitor of formula (I) or a pharmaceutically acceptable salt thereof,and ritonavir or a pharmaceutically acceptable salt thereof, in anamount effective for use as a standard or reagent in a test or assay fordetermining the ability of potential pharmaceuticals to inhibit HCVNS3/4a protease, HCV growth, or both. This aspect of the invention mayfind its use in pharmaceutical research programs.

The compounds and combinations of the present invention can be used inhigh-throughput target-analyte assays such as those for measuring theefficacy of said combination in HCV treatment.

EXAMPLES

The following examples are intended to illustrate the present inventionand not to limit it thereto.

Example 1 Preparation of Representative Intermediates Synthesis of4-hydroxy-7-methoxy-8-methyl-2-(thiazol-2-yl)quinoline (4) Step A

A solution of BCl₃ (1.0 M in CH₂Cl₂, 194 mL) was added dropwise bycanula over 20 min, under argon pressure, at 0° C. to a solution of3-methoxy-2-methylaniline (25.4 g, 185 mmol) in xylene (300 mL). Thetemperature was maintained between 0° C. and 10° C. until the additionwas completed. After an additional 30 min at 0° C., acetonitrile (12.6mL, 241 mmol) was added dropwise under argon at 0° C. After 30 min at 0°C., the resulting suspension was transferred into a dropping funnel, anddiluted with CH₂Cl₂ (40 mL). This mixture was added at 0° C. under argonover 20 min to a suspension of AlCl₃ (25.9 g, 194 mmol) in CH₂Cl₂ (40mL). The resulting orange solution was heated in an oil bath at 70° C.under a nitrogen stream for 12 h. Then, the reaction mixture was cooleddown to room temperature, and ice-cold water and CH₂Cl₂ were added. Thismixture was heated at reflux for 6 h, and then cooled to roomtemperature. After 12 h, the pH was adjusted at 0° C. to 3 with 6N NaOH.The solution was extracted with CH₂Cl₂, successively washed with water,1N NaOH, and brine. The organic layer was dried (Na₂SO₄), filtered andconcentrated in vacuo. The residue was triturated at room temperature indiisopropyl ether (50 mL) for 0.5 h. Then, the suspension was cooled at0° C., filtered, and washed with small portion of diisopropyl ether anddried under high vacuum to give 15.4 g (46%) of the desired product 2:m/z=180 (M+H)⁺.

Step B

EDCI (257 mg, 1.34 mmol) and HOAt (152 mg, 1.12 mmol) were added to astirred solution of 2 (200 mg, 1.12 mmol) in CH₂Cl₂ (10 mL) and dry DMF(1 mL). The resulting solution was stirred at room temperature for 3days. Then, the reaction mixture was partitioned between CH₂Cl₂ and 1NNaHCO₃. The organic layer was successively washed with 1N NH₄Cl, andwater, dried (Na₂SO₄), and evaporated. Purification by flashchromatography (gradient AcOEt/heptane, 10:90 to 50:50) afforded 62 mg(19%) of the target product: m/z=291 (M+H)⁺.

Step C

tBuOK (50 mg, 0.448 mmol) was added to a suspension of acetophenone 3(62 mg, 0.213 mmol) in tBuOH (5 mL). The resulting mixture was stirredat 80° C. overnight, then cooled at room temperature. The reactionmixture was diluted with AcOEt, acidified with KHSO₄, and successivelywashed with water and brine. Organic layer was dried (Na₂SO₄) andevaporated to give 43 mg (74%) of the target product as a white powder:m/z=273 (M+H)⁺.

Synthesis of (hex-5-enyl)(methyl)amine (21)

Step A

Sodium hydride (1.05 eq) was slowly added at 0° C. to a solution ofN-methyltrifluoro-acetamide (25 g) in DMF (140 mL). The mixture wasstirred for 1 h at room temperature under nitrogen. Then, a solution ofbromohexene (32.1 g) in DMF (25 mL) was added dropwise and the mixturewas heated to 70° C. for 12 hours. The reaction mixture was poured onwater (200 mL) and extracted with ether (4×50 mL), dried (MgSO₄),filtered and evaporated to give 35 g of the target product 20 as ayellowish oil which was used without further purification in the nextstep.

Step B

A solution of potassium hydroxide (187.7 g) in water (130 mL) was addeddropwise to a solution of 20 (35 g) in methanol (200 mL). The mixturewas stirred at room temperature for 12 hours. Then, the reaction mixturewas poured on water (100 mL) and extracted with ether (4×50 mL), dried(MgSO₄), filtered and the ether was distilled under atmosphericpressure. The resulting oil was purified by distillation under vacuum(13 mm Hg pressure, 50° C.) to give 7.4 g (34%) of the title product 21as a colourless oil: ¹H-NMR (CDCl₃): δ 5.8 (m, 1H), 5 (ddd, J=17.2 Hz,3.5 Hz, 1.8 Hz, 1H), 4.95 (m, 1H), 2.5 (t, J=7.0 Hz, 2H), 2.43 (s, 3H),2.08 (q, J=7.0 Hz, 2H), 1.4 (m, 4H), 1.3 (br s, 1H).

Example 2 Preparation of17-[7-methoxy-8-methyl-2-(thiazol-2-yl)quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (29) Step A

3-Oxo-2-oxa-bicyclo[2.2.1]heptane-5-carboxylic acid 22 (500 mg, 3.2mmol) in 4 ml DMF was added at 0° C. to HATU (1.34 g, 3.52 mmol) andN-methylhex-5-enylamine (435 mg, 3.84 mmol) in DMF (3 mL), followed byDIPEA. After stirring for 40 min at 0° C., the mixture was stirred atroom temperature for 5 h. Then, the solvent was evaporated, the residuedissolved in EtOAc (70 mL) and washed with saturated NaHCO₃ (10 mL). Theaqueous layer was extracted with EtOAc (2×25 mL). The organic layerswere combined, washed with saturated NaCl (20 mL), dried (Na₂SO₄), andevaporated. Purification by flash chromatography (EtOAc/petroleum ether,2:1) afforded 550 mg (68%) of the target product 23 as a colorless oil:m/z=252 (M+H)⁺.

Step B

A solution of LiOH (105 mg in 4 ml of water) was added at 0° C. to thelactone amide 23. After 1 h, the conversion was completed (HPLC). Themixture was acidified to pH 2-3 with 1N HCl, extracted with AcOEt, dried(MgSO₄), evaporated, co-evaporated with toluene several times, and driedunder high vacuum overnight to give 520 mg (88%) of the target product24: m/z=270 (M+H)⁺.

Step C

The 1-(amino)-2-(vinyl)cyclopropanecarboxylic acid ethyl esterhydrochloride 25 (4.92 g, 31.7 mmol) and HATU (12.6 g, 33.2 mmol) wereadded to 24 (8.14 g, 30.2 mmol). The mixture was cooled in an ice bathunder argon, and then DMF (100 mL) and DIPEA (12.5 mL, 11.5 mmol) weresuccessively added. After 30 min at 0° C., the solution was stirred atroom temperature for an additional 3 h. Then, the reaction mixture waspartitioned between EtOAc and water, washed successively with 0.5 N HCl(20 mL) and saturated NaCl (2×20 mL), and dried (Na₂SO₄). Purificationby flash chromatography (AcOEt/CH₂Cl₂/Petroleum ether, 1:1:1) afforded7.41 g (60%) of the target product 26 as a colorless oil: m/z=407(M+H)⁺.

Step D

DIAD (218 μL, 1.11 mmol) is added at −20° C. under nitrogen atmosphereto a solution of 26 (300 mg, 0.738 mmol), quinoline 4 (420 mg, 1.03mmol) and triphenylphosphine (271 mg, 1.03 mmol) in dry THF (15 mL).Then, the reaction is warmed up to room temperature. After 1.5 h, thesolvent is evaporated and the crude product is purified by flash columnchromatography (gradient of petroleum ether/CH₂Cl₂/ether, 3:1.5:0.5 to1:1:1) to give the target product 27: m/z=661 (M+H)⁺.

Step E

A solution of 27 (200 mg, 0.30 mmol) and Hoveyda-Grubbs 1^(st)generation catalyst (18 mg, 0.030 mmol) in dried and degassed1,2-dichloroethane (300 mL) is heated at 70° C. under nitrogen for 12 h.Then, the solvent is evaporated and the residue purified by silica gelchromatography (Petroleum ether/CH₂Cl₂/Et₂O; 3:1:1) to give the targetproduct 28: m/z=633 (M+H)⁺.

Step F

A solution of LiOH (327 mg) in water (3 mL) is added to a stirredsolution of 28 (150 mg, 0.237 mmol) in THF (15 mL) and MeOH (10 mL).After 48 h, solvent is evaporated and the residue partitioned betweenwater and ether. Aqueous layer is acidified (pH=3) and extracted withAcOEt, dried (MgSO₄) and evaporated. The residue is crystallized fromether to give the target compound 29: m/z=605 (M+H)⁺.

Example 3 Preparation ofN-[17-[7-methoxy-8-methyl-2-(thiazol-2-yl)quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(30)

A mixture of 29 (85 mg, 0.14 mmol) and CDI (47 mg, 0.29 mmol) in dry THF(7 mL) is heated at reflux for 2 h under nitrogen. LCMS analysis showsone peak of the intermediate (RT=5.37). The reaction mixture is cooledto room temperature and cyclopropylsulfonamide (52 mg, 0.43 mmol) isadded. Then, DBU (50 μL, 0.33 mmol) is added and the reaction mixture isstirred at room temperature for 1 h, and then heated at 55° C. for 24 h.Solvent is evaporated, and the residue partitioned between AcOEt andacidic water (pH=3). The crude material is purified by columnchromatography (AcOEt/CH₂Cl₂/Petroleum ether, 1:1:1). The residue iscrystallized in Et₂O, filtered to give the target compound contaminatedwith the cyclopropyl-sulfonamide. This material is triturated in 3 ml ofwater, filtered, washed with water and dried overnight in the highvacuum pump to give the target compound 30 as a white powder: m/z=708(M+H)⁺.

Example 4 Preparation of17-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (46) Synthesis of4-hydroxy-2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinoline (36)Step 1: synthesis of N-(tert-butyloxycarbonyl)-3-methoxy-2-methylaniline(32)

Triethylamine (42.4 mL, 302 mmol) was added to a suspension of3-methoxy-2-methylbenzoic acid (45.6 g, 274 mmol) in dry toluene (800mL). A clear solution was obtained. Then, dppa (65.4 mL, 302 mmol) intoluene (100 mL) was slowly added. After 1 h at room temperature, thereaction mixture was successively heated at 50° C. for 0.5 h, at 70° C.for 0.5 h then at 100° C. for 1 h. To this solution, t-BuOH (30.5 g, 411mmol) in toluene (40 mL) was added at 100° C. and the resulting mixturewas refluxed for 7 h. The solution was cooled to room temperature thensuccessively washed with water, 0.5 N HCl, 0.5 N NaOH and brine, dried(Na₂SO₄), and evaporated to give 67 g of the target product: m/z=237(M)⁺.

Step 2: synthesis of 3-methoxy-2-methylaniline (33)

TFA (40.7 mL, 548 mmol) was added to a solution ofN-(tert-butyloxycarbonyl)-3-methoxy-2-methylaniline, in dichloromethane(500 mL). After 2 h at room temperature, TFA (40.7 mL, 548 mmol) wasadded and the resulting mixture was stirred at room temperatureovernight. Then, volatiles were evaporated. The residue was trituratedwith toluene (100 mL) and diisopropylether (250 mL), filtered off andwashed with diisopropyl ether (100 mL) to give 56.3 g of the titleproduct as a TFA salt: m/z=138 (M+H)⁺. The TFA salt was transformed tothe free aniline by treatment with NaHCO₃.

Step 3: synthesis of (2-amino-4-methoxy-3-methylphenyl)(methyl)ketone(34)

A solution of BCl₃ (1.0 M, 200 mL, 200 mmol) in CH₂Cl₂ was slowly addedunder nitrogen to a solution of 3-methoxy-2-methylaniline (26.0 g, 190mmol) in xylene (400 mL). The temperature was monitored during theaddition and was kept below 10° C. The reaction mixture was stirred at5° C. for 0.5 h. Then, dry acetonitrile (13 mL, 246 mmol) was added at5° C. After 0.5 h at 5° C., the solution was transferred into a droppingfunnel and slowly added at 5° C. to a suspension of AlCl₃ (26.7 g, 200mmol) in CH₂Cl₂ (150 mL). After 45 min at 5° C., the reaction mixturewas heated at 70° C. under a nitrogen stream. After evaporation ofCH₂Cl₂, the temperature of the reaction mixture reached 65° C. After 12h at 65° C., the reaction mixture was cooled at 0° C., poured onto ice(300 g), and slowly heated to reflux for 7 h. After 2 days at roomtemperature, 6 N NaOH (50 mL) was added. The pH of the resultingsolution was 2-3. The xylene layer was decanted. The organic layer wasextracted with CH₂Cl₂. The xylene and CH₂Cl₂ layers were combined,successively washed with water, 1N NaOH, and brine, dried (Na₂SO₄) andevaporated. The residue was triturated in diisopropyl ether at 0° C.,filtered off and washed with diisopropylether to give 13.6 g (40%) ofthe title product as a yellowish solid: m/z=180 (M+H)⁺.

Step 4: synthesis of2′-[[(4-isopropylthiazole-2-yl)(oxo)methyl]amino]-4′-methoxy-3′-methylacetophenone(35)

A solution of (2-amino-4-methoxy-3-methylphenyl)(methyl)ketone (18.6 g,104 mmol) in dioxane (50 mL) was added under nitrogen to a suspension of4-isopropylthiazole-2-carbonyl chloride in dioxane (250 mL). After 2 hat room temperature, the reaction mixture was concentrated to dryness.Then, the residue was partitioned between an aqueous solution of NaHCO₃and AcOEt, organic layer was washed with brine, dried (Na₂SO₄), andevaporated. The residue was triturated in diisopropyl ether, filteredoff and washed with diisopropyl ether to give 30.8 g (90%) of the titleproduct 35.

Step 5: synthesis of4-hydroxy-2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinoline (36)

Potassium tert-butoxide (21.8 g, 195 mmol) was added to a suspension of2′-[[(4-iso-propylthiazole-2-yl)(oxo)methyl]amino]-4′-methoxy-3′-methylacetophenone(35, 30.8 g, 92.7 mmol) in tert-butanol. The resulting reaction mixtureswas heated at 100° C. overnight. Then, the reaction mixture was cooledat room temperature and diluted with ether (100 mL). The precipitate wasfiltered off and washed with Et₂O to give a powder (fraction A). Themother liquor was concentrated in vacuo, triturated in ether, filteredoff, and washed with ether to give a powder (fraction 2). Fractions 1and 2 were mixed and poured into water (250 mL). The pH of the resultingsolution was adjusted to 6-7 (control with pH paper) with HCl 1N. Theprecipitate was filtered off, washed with water and dried. Then, thesolid was triturated in diisopropyl ether, filtered off and dried togive 26 g (88%) of the title product 36 as a brownish solid: m/z=315(M+H)⁺.

Synthesis of (hex-5-enyl)(methyl)amine (38)

Step A

Sodium hydride (1.05 eq) was slowly added at 0° C. to a solution ofN-methyltrifluoro-acetamide (25 g) in DMF (140 mL). The mixture wasstirred for 1 h at room temperature under nitrogen. Then, a solution ofbromohexene (32.1 g) in DMF (25 mL) was added dropwise and the mixturewas heated to 70° C. for 12 hours. The reaction mixture was poured onwater (200 mL) and extracted with ether (4×50 mL), dried (MgSO₄),filtered and evaporated to give 35 g of the target product 37 as ayellowish oil which was used without further purification in the nextstep.

Step B

A solution of potassium hydroxide (187.7 g) in water (130 mL) was addeddropwise to a solution of 37 (35 g) in methanol (200 mL). The mixturewas stirred at room temperature for 12 hours. Then, the reaction mixturewas poured on water (100 mL) and extracted with ether (4×50 mL), dried(MgSO₄), filtered and the ether was distilled under atmosphericpressure. The resulting oil was purified by distillation under vacuum(13 mm Hg pressure, 50° C.) to give 7.4 g (34%) of the title product 38as a colourless oil: ¹H-NMR (CDCl₃): δ 5.8 (m, 1H), 5 (ddd, J=17.2 Hz,3.5 Hz, 1.8 Hz, 1H), 4.95 (m, 1H), 2.5 (t, J=7.0 Hz, 2H), 2.43 (s, 3H),2.08 (q, J=7.0 Hz, 2H), 1.4 (m, 4H), 1.3 (br s, 1H).

Preparation of17-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-3-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (46) Step A

3-Oxo-2-oxa-bicyclo[2.2.1]heptane-5-carboxylic acid 39 (500 mg, 3.2mmol) in 4 ml DMF was added at 0° C. to HATU (1.34 g, 3.52 mmol) andN-methylhex-5-enylamine (435 mg, 3.84 mmol) in DMF (3 mL), followed byDIPEA. After stirring for 40 min at 0° C., the mixture was stirred atroom temperature for 5 h. Then, the solvent was evaporated, the residuedissolved in EtOAc (70 mL) and washed with saturated NaHCO₃ (10 mL). Theaqueous layer was extracted with EtOAc (2×25 mL). The organic phaseswere combined, washed with saturated NaCl (20 mL), dried (Na₂SO₄), andevaporated. Purification by flash chromatography (EtOAc/petroleum ether,2:1) afforded 550 mg (68%) of the target product 40 as a colorless oil:m/z=252 (M+H)⁺.

Step B

A solution of LiOH (105 mg in 4 ml of water) was added at 0° C. to thelactone amide 40. After 1 h, the conversion was completed (HPLC). Themixture was acidified to pH 2-3 with 1N HCl, extracted with AcOEt, dried(MgSO₄), evaporated, co-evaporated with toluene several times, and driedunder high vacuum overnight to give 520 mg (88%) of the target product41: m/z=270 (M+H)⁺.

Step C

The 1-(amino)-2-(vinyl)cyclopropanecarboxylic acid ethyl esterhydrochloride 42 (4.92 g, 31.7 mmol) and HATU (12.6 g, 33.2 mmol) wereadded to 41 (8.14 g, 30.2 mmol). The mixture was cooled in an ice bathunder argon, and then DMF (100 mL) and DIPEA (12.5 mL, 11.5 mmol) weresuccessively added. After 30 min at 0° C., the solution was stirred atroom temperature for an additional 3 h. Then, the reaction mixture waspartitioned between EtOAc and water, washed successively with 0.5 N HCl(20 mL) and saturated NaCl (2×20 mL), and dried (Na₂SO₄). Purificationby flash chromatography (AcOEt/CH₂Cl₂/Petroleum ether, 1:1:1) afforded7.41 g (60%) of the target product 43 as a colorless oil: m/z=407(M+H)⁺.

Step D

DIAD (1.02 mL, 5.17 mmol) was added at −15° C. under nitrogen atmosphereto a solution of 43 (1.5 g, 3.69 mmol), quinoline 36 (1.39 g, 4.43 mmol)and triphenyl-phosphine (1.26 g, 4.80 mmol) in dry THF (40 mL). After4.5 h, at −15° C., the reaction mixture was partitioned between ice-coldwater and AcOEt, dried (Na₂SO₄) and evaporated. The crude material waspurified by flash column chromatography (gradient of petroleumAcOEt/CH₂Cl₂, 1:9 to 2:8) to give 1.45 g (56%) of the target product 44:m/z=703 (M+H)⁺.

Step E

A solution of 44 (1.07 g, 1.524 mmol) and Hoveyda-Grubbs 1^(st)generation catalyst (33 mg, 0.03 eq) in dried and degassed1,2-dichloroethane (900 mL) was heated at 75° C. under nitrogen for 12h. Then, the solvent was evaporated and the residue purified by silicagel chromatography (25% EtOAc in CH₂Cl₂). 620 mg (60%) of puremacrocycle 45 were obtained. m/z=674 (M+H)⁺. ¹H NMR (CDCl₃): 1.18-1.39(m, 12H), 1.59 (m, 1H), 1.70-2.08 (m, 5H), 2.28 (m, 1H), 2.38 (m, 1H),2.62 (m, 2H), 2.68 (s, 3H), 2.83 (m, 1H), 3.06 (s, 3H), 3.19 (sept,J=6.7 Hz, 1H), 3.36 (m, 1H), 3.83 (m, 1H), 3.97 (s, 3H), 4.09 (m, 2H),4.65 (td, J=4 Hz, 14 Hz, 1H), 5.19 (dd, J=4 Hz, 10 Hz, 1H), 5.31 (m,1H), 5.65 (td, J=4 Hz, 8 Hz, 1H), 7.00 (s, 1H), 7.18 (s, 1H), 7.46 (d,J=9 Hz, 1H), 7.48 (s, 1H), 8.03 (d, J=9 Hz, 1H).

Step F

A solution of lithium hydroxide (1.65 g, 38.53 mmol) in water (15 mL)was added to a stirred solution of ester 45 (620 mg, 0.920 mmol) in THF(30 mL) and MeOH (20 mL). After 16 h at room temperature, the reactionmixture was quenched with NH₄Cl sat., concentrated under reducedpressure, acidified to pH 3 with HCl 1N and extracted with CH₂Cl₂, dried(MgSO₄) and evaporated to give 560 mg (88%) of carboxylic acid 46.m/z=647 (M+H)⁺. ¹H NMR (CDCl₃): 1.11-1.40 (m, 8H), 1.42-1.57 (m, 2H),1.74 (m, 2H), 1.88-2.00 (m, 2H), 2.13 (m, 1H), 2.28 (m, 1H), 2.40 (m,1H), 2.59 (m, 2H), 2.67 (s, 3H), 2.81 (m, 1H), 2.97 (s, 3H), 3.19 (m,1H), 3.31 (m, 1H), 3.71 (m, 1H), 3.96 (s, 3H), 4.56 (dt, J=4 Hz, 12 Hz,1H), 5.23 (m, 2H), 5.66 (m, 1H), 7.01 (s, 1H), 7.10 (s, 1H), 7.22 (d,J=10 Hz, 1H), 7.45 (s, 1H), 8.00 (d, J=10 Hz, 1H).

Step G

A solution of17-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.04,6]octadec-7-ene-4-carboxylicacid 46 (138.3 mg, 0.214 mmol) prepared according to the proceduredescribed above, and carbonyldiimidazole (96.9 mg, 0.598 mmol) in dryTHF (5 mL) was stirred at reflux under nitrogen for 2 h. The reactionmixture was cooled down at room temperature and concentrated underreduced pressure. The residue was partitioned between EtOAc and HCl 1 N,the organic layer was washed with brine, dried (Na2SO₄) and evaporated.Then the solid was triturated in i-Pr ether to get 46′ as a whitepowder: m/z=629 (M+H)⁺. ¹H NMR (CDCl₃): 0.99-1.00 (m, 1H), 1.20-1.35 (m,2H), 1.39 (d, J=6.9 Hz, 6H), 1.55-1.7 (m, 1H), 1.9-2 (m, 2H), 2.15-2.25(m, 2H), 2.3-2.60 (m, 4H), 2.68 (s, 3H), 2.71-2.82 (m, 1H), 2.82-2.9 (m,1H), 3.08 (s, 3H), 3.1-3.2 (m, 1H), 3.4-3.5 (m, 1H), 3.65-3.71 (m, 1H),3.91 (s, 3H), 4.28-4.4 (m, 1H), 5.32-5.46 (m, 2H), 5.85-5.95 (m, 1H),7.00 (s, 1H), 7.22 (d, J=9.2 Hz, 1H), 7.45 (s, 1H), 8.09 (d, J=9.2 Hz,1H).

Example 5 Preparation ofN-[17-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(47)

A solution of17-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.04,6]octadec-7-ene-4-carboxylicacid 46 (560 mg, 0.867 mmol) prepared according to Example 4, andcarbonyldiimidazole (308 mg, 1.90 mmol) in dry THF (10 mL) was stirredat reflux under nitrogen for 2 h. The reaction mixture was cooled toroom temperature and cyclopropylsulfonamide (400 mg, 3.301 mmol) and DBU(286 mg, 1.881 mmol) were added. This solution was heated at 50° C. for15 h. Then, the reaction mixture was cooled down at room temperature andconcentrated under reduced pressure. The residue was partitioned betweenCH₂Cl₂ and HCl 1 N, the organic layer was washed with brine, dried(MgSO₄) and evaporated. Purification by flash chromatography (gradientof EtOAc (0 to 25%) in CH₂Cl₂) afforded 314 mg of an off-white solidwhich was further washed with water, then isopropylether, and dried inthe vacuum oven to deliver 282 mg (40%) of the pure title product 47 asa white powder: m/z=750 (M+H)⁺. ¹H NMR (CDCl₃): 0.99-1.52 (m, 14H),1.64-2.05 (m, 4H), 2.77 (m, 1H), 2.41 (m, 2H), 2.59 (m, 2H), 2.69 (s,3H), 2.92 (m, 2H), 3.04 (s, 3H), 3.19 (m, 1H), 3.40 (m, 2H), 3.98 (s,3H), 4.60 (t, J=13 Hz, 1H), 5.04 (t, J=11 Hz, 1H), 5.37 (m, 1H), 5.66(m, 1H), 6.21 (s, 1H), 7.02 (s, 1H), 7.22 (d, J=10 Hz, 1H), 7.45 (s,1H), 7.99 (d, J=10 Hz, 1H), 10.82 (broad s, 1H).

Example 6 Preparation ofN-[17-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl](1-methylcyclopropyl)sulfonamide(48)

A solution of carboxylic acid 46 (240 mg, 0.38 mmol) andcarbonyldiimidazole (2 eq) in dry THF (5 mL) was stirred at reflux undernitrogen for 2 h. The reaction mixture was cooled to room temperatureand 1-methylcyclopropylsulfonamide (2 eq) and DBU (2 eq) were added.This solution was heated at 50° C. for 15 h. Then, the reaction mixturewas cooled down to room temperature and concentrated under reducedpressure. The residue was partitioned between CH₂Cl₂ and HCl 1N, theorganic layer was washed with brine, dried (MgSO₄) and evaporated.Purification by flash chromatography (gradient of EtOAc (0 to 25%) inCH₂Cl₂) afforded 170 mg (58%) of the title compound 48 as an off-whitesolid which was further washed with water, then isopropylether, anddried in the vacuum oven: m/z=764 (M+H)⁺. ¹H NMR (acetone-d6): 0.86 (m,2H), 1.15-1.78 (m, 19H), 1.87 (m, 2H), 2.13-2.54 (m, 3H), 2.57-2.71 (m,4H), 2.96-3.25 (m, 4H), 3.54 (m, 2H), 4.02 (s, 3H), 4.58 (t, J=13 Hz,1H), 5.04 (m, 1H), 5.46 (m, 1H), 5.62 (m, 1H), 7.31 (s, 1H), 7.43 (d,J=9 Hz, 1H), 7.58 (s, 1H), 8.07 (d, J=13 Hz, 1H), 8.19 (broad s, 1H),11.44 (broad s, 1H).

Example 7 Preparation of17-[8-chloro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (25) Step A: Synthesis of(2-amino-3-chloro-4-methoxyphenyl)(methyl)ketone (50)

A solution of BCl₃ (1.0 M, 138 mL, 138 mmol) in CH₂Cl₂ was slowly addedunder nitrogen to a solution of 2-chloro-3-methoxyaniline 49 (20.6 g,131 mmol) in xylene (225 mL). The temperature was monitored during theaddition and was kept below 10° C. The reaction mixture was stirred at5° C. for 0.5 h. Then, dry acetonitrile (9.0 mL, 170 mmol) was added at5° C. After 0.5 h at 5° C., the solution was transferred into a droppingfunnel and slowly added at 5° C. to a suspension of AlCl₃ (18.4 g, 138mmol) in CH₂Cl₂ (80 mL). After 45 min at 5° C., the reaction mixture washeated at 70° C. under a nitrogen stream. After evaporation of CH₂Cl₂,the temperature of the reaction mixture reached 65° C. After 12 h at 65°C., the reaction mixture was cooled to 0° C., poured onto ice (200 g),and slowly heated to reflux for 7 h. After 2 days at room temperature, 6N NaOH (25 mL) and CH₂Cl₂ (100 mL) were added. The mixture was filtered,the filtered washed with CH₂Cl₂. The organic layer was decanted, andsuccessively washed with water, 1N NaOH, and brine, dried (Na₂SO₄) andevaporated. The residue was triturated in diisopropyl ether at 0° C.,filtered off and washed with diisopropylether to give 19.0 g (73%) ofthe title product 50 as a white solid: m/z=200 (M+H)⁺.

Step B: Synthesis of2′-[[(4-isopropylthiazole-2-yl)(oxo)methyl]amino]-3′-chloro-4′-methoxyacetophenone(51)

The title product 51 was prepared (79%) from(2-amino-3-chloro-4-methoxyphenyl)-(methyl)ketone (50) following theprocedure reported for2′-[[(4-isopropylthiazole-2-yl)(oxo)methyl]amino]-4′-methoxy-3′-methylacetophenone(35): m/z=353 (M+H)⁺.

Step C: synthesis of8-chloro-4-hydroxy-2-(4-isopropylthiazole-2-yl)-7-methoxy-quinoline (52)

The title product 52 was prepared (58%) from2′-[[(4-isopropylthiazole-2-yl)(oxo)-methyl]amino]-3′-chloro-4′-methoxyacetophenone(51) following the procedure reported for4-hydroxy-2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinoline (36):m/z=335 (M+H)⁺.

Step D: Preparation of compound 53

Compound 53 was prepared from alcohol 43 and8-chloro-4-hydroxy-2-(4-isopropyl-thiazole-2-yl)-7-methoxy-quinoline(52) following the procedure described for 44: m/z=723 (M+H)⁺.

Step E: Preparation of Compound 54

Compound 54 was prepared from 53 following the procedure described for45: m/z=695 (M+H)⁺.

Step F: Preparation of Compound 55

A solution of lithium hydroxide (3.85 g, 90.1 mmol) in water (30 mL) wasadded to a stirred solution of ester 54 (1.64 g, 2.36 mmol) in THF (55mL) and MeOH (40 mL). After 16 h at room temperature, more LiOH (1.0 g)was added. After 20 h at room temperature, the reaction mixture wasquenched with a saturated solution of NH₄Cl, concentrated under reducedpressure, acidified to pH 5 with HCl 1N, extracted with EtOAc, dried(MgSO₄) and evaporated to give 1.37 g (87%) of the carboxylic acid 55.m/z=667 (M+H)⁺.

Example 8 Preparation ofN-[17-[8-chloro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(56)

A solution of carboxylic acid 55 (1.37 g, 2.52 mmol) andcarbonyldiimidazole (2 eq) in dry THF (75 mL) was stirred at refluxunder nitrogen for 2 h. The reaction mixture was cooled to roomtemperature and cyclopropylsulfonamide (2 eq) and DBU (2 eq) were added.This solution was heated at 50° C. for 36 h. Then, the reaction mixturewas cooled down to room temperature and concentrated under reducedpressure. The residue was partitioned between EtOAc and HCl 1N, theorganic layer was washed with brine, dried (MgSO₄) and evaporated.Purification by flash chromatography (gradient of EtOAc (0 to 25%) inCH₂Cl₂) afforded 880 mg (55%) of the title compound 56 as an off-whitesolid: m/z=770 (M+H)⁺. ¹H NMR (CDCl₃, major rotamer): 0.93-1.52 (m,13H), 1.60-2.07 (m, 5H), 2.21-2.64 (m, 5H), 2.92 (m, 2H), 3.04 (s, 3H),3.19 (m, 1H), 3.41 (m, 2H), 4.07 (s, 3H), 4.60 (t, J=13 Hz, 1H), 5.04(t, J=11 Hz, 1H), 5.37 (m, 1H), 5.66 (m, 1H), 6.33 (s, 1H), 7.07 (s,1H), 7.24 (d, J=9 Hz, 1H), 7.52 (s, 1H), 8.05 (d, J=9 Hz, 1H), 10.81(broad s, 1H).

Example 9 Preparation ofN-[17-[8-chloro-2-(4-isopropylthiazole-2-yl)-7-methoxy-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl](1-methylcyclopropyl)sulfonamide (57)

A solution of carboxylic acid 55 (49 mg, 0.073 mmol) andcarbonyldiimidazole (2 eq) in dry THF (5 mL) was stirred at reflux undernitrogen for 2 h. The reaction mixture was cooled to room temperatureand 1-methylcyclopropylsulfonamide (2 eq) and DBU (2 eq) were added.This solution was heated at 50° C. for 15 h. Then, the reaction mixturewas cooled down to room temperature and concentrated under reducedpressure. The residue was partitioned between EtOAc and HCl 1N, theorganic layer was washed with brine, dried (MgSO₄) and evaporated.Purification by flash chromatography (gradient of EtOAc (0 to 25%) inDCM) afforded 10 mg (20%) of the title compound 57: m/z=784 (M+H)⁺.

Example 10 Preparation of17-[2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (65) Step 1: Synthesis of ethyl4-hydroxy-7-methoxy-8-methylquinoline-3-carboxylate (58)

Diethyl ethoxymethylenemalonate (17.2 g, 79.6 mmol) was added to2-methyl-m-anisidine (8.4 g, 61.2 mmol) (exothermic reaction). Then,diethylether (100 mL) was added and the mixture was stirred overnight atroom temperature. The solvent was evaporated and the residuere-dissolved in ether (50 mL), filtered, washed with heptane and driedto give 12 g of an intermediate. This intermediate was added portionwise to diphenyl ether (50 mL) pre-heated at 230° C. The reactionmixture was successively heated to 250° C. for 1.5 h, cooled at roomtemperature, and diluted with heptane (200 mL). The precipitate wasfiltered off, and successively washed with heptane and ether to give 9.2g (57.5%) of the target product 58 as a yellow powder: m/z=262 (M+H)⁺.

Step 2: Synthesis of 4-Hydroxy-7-methoxy-8-methylquinoline (59)

A suspension of ethyl4-hydroxy-7-methoxy-8-methylquinoline-3-carboxylate (58, 9.2 g, 35.2mmol) in 5N NaOH (150 mL) was refluxed for 1.5 h (until a clear solutionwas obtained). Then, the solution was cooled to 0° C. and the pHadjusted to 2-3 with concentrated HCl. The solid was filtered off andsuccessively washed with water, acetone and ether. This powder was addedin small portions to diphenylether (40 mL), pre-heated at 250° C. Theresulting suspension became a solution after 20 min (CO₂ formation wasobserved). After 1 h at 250° C., the brown solution was cooled to roomtemperature and diluted with heptanes (200 mL). The precipitate wasfiltered off and washed with heptanes and ether to give 6.4 g (96%) ofthe target product 59 as a yellow powder: m/z=190 (M+H)⁺.

Step 3: Synthesis of 4-Chloro-7-methoxy-8-methylquinoline (60)

A solution of 4-hydroxy-7-methoxy-8-methylquinoline (59, 6.4 g, 33.8mmol) in POCl₃ (17.2 g, 111.6 mmol) was heated at reflux for 1 h undernitrogen. Then, the resulting solution was cooled down to roomtemperature and the excess of POCl₃ was evaporated under reducedpressure. The residue was partitioned between ice-cold 1N NaOH andAcOEt. The organic layer was dried (Na₂SO₄), and evaporated. The residuewas purified by silica-gel filtration (AcOEt/CH₂Cl₂/Heptane, 4:4:2) togive 6.5 g (92.5%) of the target product 60 as yellow needles: m/z=208(M+H)⁺.

Step 4: Synthesis of 4-Chloro-7-methoxy-8-methylquinoline N-oxide (61)

Metachloroperbenzoic acid (90.2 g, 366.0 mmol) was added portion wiseover 3 h to a solution of 4-chloro-7-methoxy-8-methylquinoline (60, 15.2g, 73.2 mmol) in CHCl₃ (1 L). Then, the solution was partitioned betweenice-cooled NaOH 1N and CH₂Cl₂ (8 successive extractions). The organiclayers were combined, dried (Na₂SO₄) and evaporated. The residue waspurified by column chromatography (gradient of AcOEt/CH₂Cl₂, 1:2 to 1:0)to give 3.0 g (18.3%) of the title product 61 as a pale yellow powder:m/z=224 (M+H)⁺.

Step 5: Synthesis of 4-Benzyloxy-7-methoxy-8-methylquinoline N-oxide(62)

NaH (973 mg, 60% in mineral oil, 24.3 mmol) was added at 0° C., underinert atmosphere, to benzylalcohol (2.96 mL, 28.6 mmol) in DMF (10 mL).After 5 min at 0° C., the solution was warmed up to room temperature.After 10 min at room temperature, 4-chloro-7-methoxy-8-methylquinolineN-oxide (61, 3.2 g, 14.3 mmol) was added in one potion. The resultingblack solution was stirred at room temperature under inert atmospherefor another 30 min, then poured into ice-cooled water, and extracted 4times with AcOEt. Combined organic layers were dried (Na₂SO₄) andevaporated. The residue was purified by column chromatography (gradientAcOEt/CH₂Cl₂, 1:1 to 1:0, then AcOEt/MeOH 9:1) to give 2.5 g (59%) ofthe target product 62 as a yellow powder: m/z=296 (M+H)⁺.

Step 6: Synthesis of 4-benzyloxy-2-chloro-7-methoxy-8-methylquinoline(63)

POCl₃ was added under inert atmosphere at −78° C. to4-benzyloxy-7-methoxy-8-methylquinoline N-oxide (62, 2.5 g, 8.47 mmol).Then the reaction mixture was allowed to warm up to room temperature,then heated to reflux. After 35 min, the solution was cooled to roomtemperature and the excess of POCl₃ was evaporated under reducedpressure. The residue was partitioned between ice-cooled water andAcOEt, dried (Na₂SO₄) and evaporated. The residue was triturated inether, then filtered and successively washed with small portions ofmethanol and ether to give 2.4 g (90.4%) of the target product 63 as awhite powder: m/z=314 (M+H)⁺.

Step 7: Synthesis of4-hydroxy-2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methyl-quinoline (64)

A mixture of 4-benzyloxy-2-chloro-7-methoxy-8-methylquinoline (63, 1.00g, 3.19 mmol) and 3-isopropylpyrazole was heated at 155° C. for 12 h.Then, the reaction mixture was partitioned between AcOEt and water,dried (Na₂SO₄) and evaporated. The residue was purified by columnchromatography (AcOEt/CH₂Cl₂, 1:1) to give 900 mg (95%) of the targetproduct 64 as a yellowish powder: m/z=298 (M+H)⁺.

Step 8: Synthesis of17-[2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yl-oxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (65)

The title compound was prepared from4-hydroxy-2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methylquinoline (64)and intermediate 26 following the procedure (Step D-F) reported for thepreparation of17-[7-methoxy-8-methyl-2-(thiazol-2-yl)quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (29): m/z=630 (M+H)⁺.

Example 11 Preparation ofN-[17-[2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(66)

The title compound was prepared from17-[2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (65) and cyclopropylsulfonamide following the procedure reportedfor the preparation ofN-[17-[8-chloro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(56): m/z=733 (M+H)⁺. ¹H NMR (CDCl₃): 0.80-1.50 (m, 12H), 1.65-1.78 (m,1H), 1.79-2.05 (m, 4H), 2.15-2.31 (m, 1H), 2.32-2.48 (m, 2H), 2.49-2.63(m, 5H), 2.84-2.96 (m, 2H), 3.03 (s, 3H), 3.05-3.14 (m, 1H), 3.33-3.42(m, 2H), 3.61-3.70 (m, 1H), 3.96 (s, 3H), 4.60 (t, J=12.3 Hz, 1H), 5.04(t, J=10.6 Hz, 1H), 5.26-5.46 (m, 1H), 5.61-5.69 (m, 1H), 6.32 (d, J=2.5Hz, 1H), 6.37 (br s, 1H), 7.13 (d, J=9.0 Hz, 1H), 7.30 (s, 1H), 7.95 (d,J=9.0 Hz, 1H), 8.68 (d, J=2.5 Hz, 1H), 10.88 (br s, 1H).

Example 12 Preparation of17-[8-ethyl-2-(4-isopropylthiazole-2-yl)-7-methoxy-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (70) Step 1: Synthesis ofN-[2-(1-hydroxyethyl)-3-methoxyphenyl]pivaloylamide (66)

A solution of N-butyllithium (2.5 M in hexanes, 4.4 mL, 11.1 mmol) wasadded dropwise at 0° C. under nitrogen to a stirred solution ofN-(3-methoxyphenyl)-pivaloylamide. After 1 h at room temperature, thereaction mixture was cooled down to −78° C. Then, a solution ofacetaldehyde (544 μL, 9.64 mmol) in THF (1 mL) was added. After 10 min,the reaction mixture was allowed to warm up to room temperature for 30min. Then, the reaction mixture was partitioned between AcOEt and water,dried (Na₂SO₄) and evaporated to afford 500 mg (45%) of the targetproduct 66 as a yellow solid: m/z=252 (M+H)⁺.

Step 2: Synthesis of N-[2-ethyl-3-methoxyphenyl]pivaloylamide (67)

A mixture of N-[2-(1-hydroxyethyl)-3-methoxyphenyl]pivaloylamide (66, 42g, 167 mmol), Pd/C (10%, 2.00 g) and H₂SO₄ (10 mL) in acetic acid (400mL) was stirred at room temperature for 30 minutes. Then, the resultingreaction mixture was hydrogenated for 4 days, after which the catalystwas eliminated by filtration on kieselghur. The filtrate wasconcentrated to 300 mL, then poured into 1.0 L of water. The solidformed was filtered off, washed with water to give the target product 67as a yellow solid: m/z=236 (M+H)⁺.

Step 3: Synthesis of 2-ethyl-m-anisidine (68)

A solution of N-[2-ethyl-3-methoxyphenyl]pivaloylamide (67, 167 mmol)and 37% HCl (700 mL) in EtOH (700 mL) was refluxed for 48 h. Then, thereaction mixture was cooled to room temperature and concentrated underreduced pressure (⅓ of volume). This solution was maintained at 5° C.for 6 h. The solid that appeared was filtered off, and washed withdiisopropylether to give 22.35 g of the target product as its HCl salt.The free based was generated by treatment with K₂CO₃ to give 20.85 g(83%) of the target product 68: m/z=152 (M+H)⁺.

Step 4: Synthesis of8-ethyl-4-hydroxy-2-(4-isopropylthiazole-2-yl)-7-methoxyquinoline (69)

The title compound was prepared from 2-ethyl-m-anisidine (68) followingthe procedure (Steps 3-5) reported for the preparation of4-hydroxy-2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinoline (36):m/z=329 (M+H)⁺.

Step 5: Synthesis of17-[8-ethyl-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yl-oxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (70)

The title compound was prepared from8-ethyl-4-hydroxy-2-(4-isopropylthiazole-2-yl)-7-methoxyquinoline (69)and intermediate 43 following the procedure (Steps D-F) reported for thepreparation of17-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (46): m/z=661 (M+H)⁺.

Example 13N-[17-[8-ethyl-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl](cyclo-propyl)sulfonamide(71)

The title compound was prepared from17-[8-ethyl-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carboxylicacid (70) and cyclopropylsulfonamide following the procedure reportedfor the preparation ofN-[17-[8-chloro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(56): m/z=764 (M+H)⁺.

Example 14 Preparation of17-[8-fluoro-2-(4-isopropylthiazole-2-yl)-7-methoxy-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carboxylicacid (73) Step 1:8-fluoro-4-hydroxy-2-(4-isopropylthiazole-2-yl)-7-methoxyquinoline (72)

The title compound was prepared from 2-fluoro-3-methoxybenzoic acidfollowing the procedure (steps 1-5) reported for the preparation of4-hydroxy-2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinoline (36):m/z=319 (M+H)⁺.

Step 2: Synthesis of17-[8-fluoro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (73)

The title compound was prepared from8-fluoro-4-hydroxy-2-(4-isopropylthiazole-2-yl)-7-methoxyquinoline (72)and alcohol 43 following the procedure (steps D-F) reported for thepreparation of17-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (46): m/z=651 (M+H)⁺.

Example 15N-[17-[8-fluoro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yl-oxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl]-(cyclopropyl)sulfonamide(74)

The title compound was prepared from17-[8-fluoro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carboxylicacid (73) and cyclopropylsulfonamide following the procedure reportedfor the preparation ofN-[17-[8-chloro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(56): m/z=754 (M+H)⁺. ¹H NMR (CDCl₃): ¹H NMR (CDCl₃): 0.75-1.52 (m,15H), 1.64-2.05 (m, 4H), 2.77 (m, 1H), 2.41 (m, 2H), 2.59 (m, 2H), 2.92(m, 2H), 3.04 (s, 3H), 3.19 (m, 1H), 3.40 (m, 2H), 4.07 (s, 3H), 4.60(m, 1H), 5.05 (t, J=10.5 Hz, 1H), 5.37 (m, 1H), 5.66 (m, 1H), 6.17 (s,1H), 7.07 (s, 1H), 7.54 (s, 1H), 7.86 (m, 1H), 10.77 (broad s, 1H).

Example 1618-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-2,15-dioxo-3,14-diazatricyclo[14.3.0.0^(4,6)]nonadec-7-ene-4-carboxylicacid (80) Step 1. Synthesis of N-(hept-6-enyl)phthalimide (75)

A solution of potassium phthalimide (627 mg, 3.38 mmol) and7-bromohept-1-ene in dry DMF (10 mL) was stirred at 100° C. undernitrogen for 1 h. Then, the reaction mixture was successively cooled toroom temperature, filtered, diluted with ether, and filtered again. Thefiltrate was concentrated under reduced pressure to give the targetproduct 75 as an oil, which was used without further purifications inthe next step: m/z=244 (M+H)⁺.

Step 2. Synthesis of 6-heptenylamine (76)

A solution of N-(hept-6-enyl)phthalimide (75, 66.2 g, 272 mmol) andhydrazine hydrate (19.8 mL, 408 mmol) in MeOH (1.0 L) was stirred atroom temperature overnight. Then, the reaction mixture was cooled toroom temperature and the solid discarded by filtration. The filtrate wasdiluted with ether and the solid formed discarded by filtration. Theether was evaporated under reduced pressure. Then, 5N HCl (50 mL) wasadded and the resulting mixture was stirred at reflux. After 45 min.,the reaction mixture was cooled down to room temperature and the solidformed filtered. The pH of the filtrate was adjusted to 3 at 0° C. withNaOH. Then, the reaction mixture was extracted with ether and dried(Na₂SO₄) and evaporated. The crude was purified by distillation to give34.57 g of the target product 76 as an oil: m/z=114 (M+H)⁺.

Step 3. Synthesis of intermediate 77

The title compound was prepared from 6-heptenylamine (76) and3-Oxo-2-oxa-bicyclo[2.2.1]heptane-5-carboxylic acid (22) following theprocedure reported for the preparation of intermediate 23: m/z=252(M+H)⁺. The title compound was also prepared (82% isolated yield) usingother coupling conditions (EDCI.HCl (1.1 eq.), HOAT (1.1 eq.) anddiisopropylethylamine in dry DMF).

Step 4. Synthesis of intermediate 78

The title compound was prepared (65%) from intermediate 77 and LiOHfollowing the procedure reported for the preparation of intermediate 24:m/z=270 (M+H)⁺.

Step 5. Synthesis of intermediate 79

The title compound was prepared (65%) from intermediate 78 and1-(amino)-2-(vinyl)cyclopropanecarboxylic acid ethyl ester hydrochloride25 following the procedure reported for the preparation of intermediate26: m/z=407 (M+H)⁺.

Step 6. Synthesis of18-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-2,15-dioxo-3,14-diazatricyclo[14.3.0.0^(4,6)]nonadec-7-ene-4-carboxylicacid (80)

The title compound was prepared from intermediate 79 and quinoline 36following the procedure (Steps D-F) reported for the preparation of17-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo-[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (46): m/z=647 (M+H)⁺.

Example 17N-[18-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinolin-4-yl-oxy]-2,15-dioxo-3,14-diazatricyclo[14.3.0.0^(4,6)]nonadec-7-ene-4-carbonyl](cyclo-propyl)sulfonamide(81)

The title compound was prepared from18-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-2,15-dioxo-3,14-diazatricyclo[14.3.0.0^(4,6)]nonadec-7-ene-4-carboxylicacid (80) and cyclopropylsulfonamide following the procedure reportedfor the preparation ofN-[17-[2-(4-isopropylthiazole-2-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(47): m/z=750 (M+H)⁺. ¹H NMR (CDCl₃): 0.90-0.96 (m, 1H), 1.1-1.2 (m,4H), 1.39 (d, J=6.9 Hz, 6H), 1.4-1.55 (m, 5H), 1.80-1.92 (m, 5H),2.15-2.25 (m, 1H), 2.30-2.40 (m, 1H), 2.45-2.55 (m, 2H), 2.68 (s, 3H),2.85-2.92 (m, 1H), 3.15-3.30 (m, 2H), 3.45-3.55 (m, 2H), 3.96 (s, 3H),4.09 (dd, J=11.5 Hz, J=3.8 Hz, 1H), 4.61 (t, J=7.9 Hz, 1H), 4.99 (t,J=9.0 Hz, 1H), 5.51-5.53 (m, 1H), 5.71 (dd, J=18.6 Hz, J=8.2 Hz, 1H),6.86 (s, 1H), 7.03 (s, 1H), 7.20 (d, J=9.2 Hz, 1H), 7.50 (s, 1H), 7.88(d, J=9.2 Hz, 1H), 9.40 (br s, 1H).

Example 18N-[[18-[2-[4-(isopropyl)thiazol-2-yl]-7-methoxy-8-methylquinolin-4-yloxy]-2,15-dioxo-14-(4-methoxybenzyl)-3,14,16-triazatricyclo[14.3.0.0^(4,6)]nonadec-7-en-4-yl]carbonyl](cyclopropyl)sulfonamide(90)

Step A: Synthesis of intermediate 82

Boc-cis-hydroxy-L-Proline methyl ester (500 mg, 2.04 mmol),4-hydroxy-2-[4-(iso-propyl)thiazol-2-yl]-7-methoxy-8-methylquinoline(36, 769 mg, 2.04 mmol) and 2-diphenylphosphanylpyridine (751 mg, 2.86mmol) were dried under high vacuum for 1 h. Dry THF was then added undernitrogen and the resulting reaction mixture was cooled to −15° C. Then,DIAD was added drop wise. After 1 h at −5° C. the solution was allowedto warm up to room temperature. After 16 h, the reaction mixture waspartitioned between ice-cold water and AcOEt. The organic layer wassuccessively washed vigorously with HCl 1M and brine, dried (MgSO₄),filtered and evaporated. Purification by column chromatography on silicagel (gradient AcOEt/CH₂Cl₂, 0:10 to 5:95) afforded 940 mg (85%) of thedesired product 82 as a colorless oil: m/z=542 (M+H)⁺.

Step B: Synthesis of intermediate 83

A solution of LiOH (592 mg, 13.8 mmol) in water was added to a solutionof intermediate 82 (1.5 g, 2.77 mmol) in MeOH/THF 1:1. After 16 h atroom temperature, the reaction mixture was acidified to pH 3-4 withdiluted HCl, extracted with AcOEt, washed with brine, dried (MgSO₄) andevaporated. The residue was purified by flash chromatography (GradientAcOEt/CH₂Cl₂, 1:9 to 4:6) to give 1.26 g (86%) of the title product 83as an orange oil: m/z=528 (M+H)⁺.

Step C: Synthesis of intermediate 84

To a stirred solution of carboxylic acid 83 (1.26 g, 2.39 mmol) in dryDMF (20 mL) was added (1R,2S)-1-amino-2-vinylcyclopropanecarboxylic acidethyl ester tosylate (860 mg, 2.63 mmol) and diisopropylethylamine (1.04mL, 5.98 mmol). Then, HATU (999 mg, 2.63 mmol) was added at 0° C. undernitrogen. The resulting solution was stirred at 0° C. for 30 minutes,then at room temperature. After 4 h, the reaction mixture was dilutedwith water and extracted with AcOEt. The organic layers were combinedand successively washed with a saturated solution of NaHCO₃, water andbrine, dried (MgSO₄), and evaporated. Purification by columnchromatography (gradient AcOEt/CH₂Cl₂, 0:1 to 2:8) afforded 1.44 g (90%)of the title product 84 as a white solid: m/z=665 (M+H)⁺.

Step D: Synthesis of intermediate 85

To a stirred solution of Boc-protected proline derivative 84 (1.44 g,2.16 mmol) in CH₂Cl₂ (20 mL) was added trifluoroacetic acid (5 mL).After 2 h at room temperature, the reaction mixture was concentrated andthe residue was partitioned between a saturated solution of NaHCO₃ andCH₂Cl₂. The organic layer was dried (MgSO₄) filtered and concentrated togive 1.0 g (81%) of the title product 85 as a colorless oil: m/z=565(M+H)⁺.

Step E: Synthesis of N-(hept-6-enyl)-N-(4-methoxybenzyl)amine 86

A solution of hept-6-enylamine (2.0 g, 13.4 mmol) and anisaldehyde (1.79mL, 14.7 mmol) in EtOH (50 mL) was stirred at room temperature for 1 h.Then, NaBH₄ (556 mg, 14.7 mmol) was added at 0° C. under nitrogen. Theresulting solution was allowed to warm up to room temperature for 4 h.Then, the reaction mixture was partitioned between ice-cold water andCH₂Cl₂, washed with brine, dried (Na₂SO₄) and evaporated. The residuewas purified by chromatography (gradient AcOEt/CH₂Cl₂ 0:1 to 2:8, thenCH₂Cl₂/MeOH 9:1) to give 1.8 g (34%) of the title product 86 as acolorless oil: m/z=234 (M+H)⁺.

Step F: Synthesis of intermediate 87

To a solution of proline derivative 85 in THF (50 mL) was added NaHCO₃(1.0 g). Then, phosgene (4.7 mL, 20% solution in toluene) was added at0° C. under nitrogen. After 1.5 h, the white solid was filtered off andwashed with THF and CH₂Cl₂. Then, the filtrate was concentrated underreduced pressure and the residue was re-dissolved in dry dichloromethane(50 mL). To this solution, NaHCO₃ (1.0 g) and protected amine 86 weresuccessively added. After 16 h at room temperature, the reaction mixturewas filtered off. The filtrate was concentrated under reduced pressureand the resulting residue was purified by silica chromatography(gradient AcOEt/CH₂Cl₂, 0:1 to 2:8) to give 1.36 g (90%) of the titleproduct 87: m/z=824 (M+H)⁺.

Step G: Synthesis of intermediate 88

Hoveyda-Grubbs 1^(st) generation catalyst (50 mg, 0.082 mmol) was addedto a degassed solution of diene 87 (1.36 g, 1.65 mmol) in toluene (170mL). The resulting solution was heated at 80° C. under nitrogen for 4 h.Then, the reaction mixture was concentrated and purified by flashchromatography (gradient AcOEt/CH₂Cl₂, 0:1 to 2:8) to give 900 mg (65%)of the title product 88 as a brownish foam: m/z=796 (M+H)⁺.

Step H: Synthesis of intermediate 89

A solution of LiOH (242 mg, 5.65 mmol) in water (20 mL) was added to asolution of ester 88 (900 mg, 1.13 mmol) in MeOH/THF 1:1. The reactionmixture was stirred at 50° C. for 2 h, then cooled down to roomtemperature, acidified to pH 3-4 with diluted HCl, and extracted withAcOEt. The organic layers were successively combined, washed with brine,dried (MgSO₄), filtered and evaporated to give 840 mg (97%) of the titleproduct 89 as a slightly yellow solid: m/z=768 (M+H)⁺.

Step I: Synthesis ofN-[[18-[2-[4-(isopropyl)thiazol-2-yl]-7-methoxy-8-methylquinolin-4-yloxy]-2,15-dioxo-14-(4-methoxybenzyl)-3,14,16-triazatricyclo[14.3.0.0^(4,6)]nonadec-7-en-4-yl]carbonyl](cyclopropyl)sulfonamide(90)

A solution of carboxylic acid 65 (830 mg, 1.03 mmol) andcarbonyldiimidazole (333 mg, 2.06 mmol) in dry THF (20 mL) was stirredat reflux under nitrogen for 2 h. Then, the reaction mixture was cooledto room temperature and cyclopropyl-sulfonamide (249 mg, 2.06 mmol) andDBU (313 mg, 2.06 mmol) were added. The resulting solution was stirredat 50° C. for 12 h, then cooled to room temperature. The reactionmixture was quenched with water and extracted with CH₂Cl₂, washed withdiluted HCl, dried (MgSO₄), filtered and evaporated. The crude materialwas purified by column chromatography (CH₂Cl₂/EtOAc, 80:20) andrecrystallized from CH₂Cl₂/ether to give 450 mg (50%) of the titleproduct 90 as a white powder: m/z=871 (M+H)⁺; ¹H-NMR (CDCl₃): 1.05-1.61(m, 18H), 2.00 (m, 1H), 2.12-2.22 (m, 2H), 2.59-2.70 (m, 5H), 2.96 (m,1H), 3.15-3.20 (m, 3H), 3.63 (s, 3H), 3.71-3.78 (m, 2H), 3.88-3.94 (m,4H), 4.54 (d, J=15 Hz, 1H), 5.08 (t, J=8.5 Hz, 1H), 5.16 (t, J=9.4 Hz,1H), 5.38 (m, 1H), 5.75 (m, 1H), 6.45 (d, J=8.4 Hz, 2H), 6.65 (d, J=8.4Hz, 2H), 7.03 (s, 1H), 7.10 (d, J=9.1 Hz, 1H), 7.41 (s, 1H), 7.73 (d,J=9.1 Hz, 1H), 7.76 (br s, 1H), 10.15 (br s, 1H).

Example 19N-[[18-[2-[4-(isopropyl)thiazol-2-yl]-7-methoxy-8-methylquinolin-4-yloxy]-2,15-dioxo-3,14,16-triazatricyclo[14.3.0.0^(4,6)]nonadec-7-en-4-yl]carbonyl]-(cyclopropyl)sulfonamide(91)

TFA (10 mL) was added to a solution ofN-[[18-[2-[4-(isopropyl)thiazol-2-yl]-7-methoxy-8-methylquinolin-4-yloxy]-2,15-dioxo-14-(4-methoxybenzyl)-3,14,16-triazatricyclo[14.3.0.0^(4,6)]nonadec-7-en-4-yl]carbonyl](cyclopropyl)sulfonamide(90) in DCM (20 mL). After 30 min at room temperature, water (20 mL) wasadded to the reaction mixture and the pH was adjusted to 3-4 withNaHCO₃. The organic layer was washed with brine, dried (Na₂SO₄),filtered and evaporated. The residue was purified by columnchromatography (gradient MeOH/CH₂Cl₂, 0:1 to 1:99, then AcOEt/CH₂Cl₂1:1) to afford 313 mg (73%) of the desired title product 91 as ayellowish solid: m/z=751 (M+H)⁺. ¹H-NMR (CDCl₃): 0.88-1.64 (m, 16H),1.96 (m, 2H), 2.52 (m, 1H), 2.68 (m s, 5H), 2.79-2.92 (m, 3H), 3.18 (m,1H), 3.63-3.69 (m, 2H), 3.86 (m, 1H), 3.97 (s, 3H), 4.34 (m, 1H), 4.59(m, 1H), 5.08 (m, 1H), 5.40 (m, 1H), 5.80 (m, 1H), 6.73 (s, 1H), 7.03(s, 1H), 7.21 (d, J=8.9 Hz, 1H), 7.26 (br s, 1H), 7.47 (s, 1H), 7.92 (d,J=8.9 Hz, 1H), 10.20 (br s, 1H).

Example 20N-[[18-[8-chloro-2-[4-(isopropyl)thiazol-2-yl]-7-methoxyquinolin-4-yloxy]-2,15-dioxo-3,14,16-triazatricyclo[14.3.0.0^(4,6)]nonadec-7-en-4-yl]carbonyl]-(cyclopropyl)sulfonamide(94) Step A: Synthesis of4,8-dichloro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinoline (92)

A solution of8-chloro-4-hydroxy-2-(4-isopropylthiazole-2-yl)-7-methoxy-quinoline (2.0g, 5.97 mmol) in POCl₃ (10 mL) was heated at 85° C. during 30 min. Then,the reaction mixture was concentrated under reduced pressure. Theresidue was poured into ice-cooled water (20 mL), the pH was adjusted to10 with 50% NaOH, and extracted with CH₂Cl₂. The organic layer waswashed with brine, dried (MgSO₄), filtered, and evaporated to give 2.05g (97%) of the title compound 92 as a yellow solid: m/z=353 (M+H)⁺.

Step B: Synthesis of intermediate 93

NaH (60% in mineral oil, 679 mg, 17.0 mmol) was added under nitrogen toa solution of Boc-trans-hydroxy-L-Proline-OH (2.0 g, 5.661 mmoles) indry DMF (50 mL). After 30 min at room temperature, a solution of4,8-dichloro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinoline (92, 1.38g, 5.94 mmol) in dry DMF was added and the resulting solution wasstirred overnight at room temperature. Then, the reaction mixture wasquenched with diluted HCl until pH 2, extracted twice with AcOEt, andthe combined organic layers were washed with brine, dried (MgSO₄) andevaporated. The residue was purified by column chromatography (gradientAcOEt/CH₂Cl₂, 0:1 to 1:1) to give 2.35 g (75%) of the title 93: m/z=548(M+H)⁺.

Step C: Synthesis ofN-[[18-[8-chloro-2-[4-(isopropyl)thiazol-2-yl]-7-methoxy-quinolin-4-yloxy]-2,15-dioxo-3,14,16-triazatricyclo[14.3.0.0^(4,6)]nonadec-7-en-4-yl]carbonyl](cyclopropyl)sulfonamide(94)

The title compound was synthesized from intermediate 93 following theprocedure (Steps C-I) reported forN-[[18-[2-[4-(isopropyl)thiazol-2-yl]-7-methoxy-8-methyl-quinolin-4-yloxy]-2,15-dioxo-14-(4-methoxybenzyl)-3,14,16-triazatricyclo-[14.3.0.0^(4,6)]nonadec-7-en-4-yl]carbonyl](cyclopropyl)sulfonamide(90) and forN-[[18-[2-[4-(isopropyl)thiazol-2-yl]-7-methoxy-8-methylquinolin-4-yloxy]-2,15-dioxo-3,14,16-triazatricyclo[14.3.0.0^(4,6)]nonadec-7-en-4-yl]carbonyl](cyclopropyl)-sulfonamide(91): m/z=771 (M)⁺; ¹H-NMR (CDCl₃): 0.93 (m, 1H), 1.06-1.63 (m, 15H),1.92 (m, 3H), 2.50 (m, 1H), 2.64 (m, 2H), 2.76 (m, 1H), 2.87 (m, 2H),3.20 (m, J=6.9 Hz, 1H), 3.70 (m, 1H), 3.77-3.87 (m, 1H), 4.00 (dd, J=4.0Hz, 10.1 Hz, 1H), 4.04 (s, 3H), 4.42 (m, 1H), 4.59 (t, J=7.3 Hz, 1H),5.05 (dd, J=8.3 Hz, 9.9 Hz, 1H), 5.51 (m, 1H), 5.79 (m, 1H), 7.03 (m,1H), 7.08 (s, 1H), 7.22 (d, J=9.3 Hz, 1H), 7.54 (s, 1H), 7.95 (d, J=9.3Hz, 1H).

Example 21N-[[18-[8-chloro-2-[4-(isopropyl)thiazol-2-yl]-7-methoxyquinolin-4-yloxy]-2,15-dioxo-3,14,16-triazatricyclo[14.3.0.0^(4,6)]nonadec-7-en-4-yl]carbonyl](1-methylcyclopropyl)sulfonamide(95)

The title compound was synthesized from intermediate 93 and1-Methyl-cyclopropyl-sulfonamide following the procedure (Steps C-I)reported forN-[[18-[2-[4-(isopropyl)-thiazol-2-yl]-7-methoxy-8-methylquinolin-4-yloxy]-2,15-dioxo-14-(4-methoxybenzyl)-3,14,16-triazatricyclo[14.3.0.0^(4,6)]nonadec-7-en-4-yl]carbonyl](cyclopropyl)-sulfonamide(90) and forN-[[18-[2-[4-(isopropyl)thiazol-2-yl]-7-methoxy-8-methyl-quinolin-4-yloxy]-2,15-dioxo-3,14,16-triazatricyclo[14.3.0.0^(4,6)]-nonadec-7-en-4-yl]carbonyl](cyclopropyl)sulfonamide(91): m/z=785 (M)⁺. ¹H-NMR (CDCl₃): 0.90 (m, 1H), 1.12-1.60 (m, 16H),1.74 (m, 1H), 1.90-1.99 (m, 4H), 2.51 (m, 1H), 2.65-2.78 (m, 3H), 2.88(m, 1H), 3.20 (m, J=6.7 Hz, 1H), 3.69 (m, 1H), 3.84 (m, 1H), 3.96-4.00(m, 1H), 4.01 (s, 3H), 4.46 (m, 1H), 4.63 (t, J=7.4 Hz, 1H), 5.09 (t,J=9.1 Hz, 1H), 5.50 (m, 1H), 5.79 (m, 1H), 7.08 (m, 2H), 7.22 (d, J=9.2Hz, 1H), 7.52 (s, 1H), 7.95 (d, J=9.2 Hz, 1H), 10.08 (br s, 1H).

Example 22 Cyclopropanesulfonic acid{17-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl}-amide(103)

Step A: Synthesis of 6-methylpyridine-2-carboxylic acid(6-acetyl-3-methoxy-2-methylphenyl)-amide (96)

6-Methylpicolinic acid (1.12 g, 8.167 mmol) was dissolved in dry DCM(100 ml) and kept on an ice-bath. Then,6-acetyl-3-methoxy-2-methylaniline (1.48 g, 8.17 mmol) and pyridine (6.6mL, 0.082 mol) were added followed by drop wise addition of POCl₃ (1.53mL, 0.016 mol) over 15 minutes. The resulting solution was stirred at−5° C. for 1 h. Then, water (100 mL) was added carefully and after 5 minof stirring, NaOH (40%, 20 mL) was subsequently added drop wise followedby the separation of the organic layer. The water layer was extractedthree times with CH₂Cl₂, and the combined organic layers were washedwith brine, dried (MgSO₄), filtered and evaporated. The residue waspurified by column chromatography (Heptane/AcOEt, 3:1) to give the titlecompound (2.1 g, 86%): m/z=299 (M+H)⁺.

Step B: Synthesis of4-hydroxy-2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinoline (97)

To a solution of 6-methylpyridine-2-carboxylic acid(6-acetyl-3-methoxy-2-methyl-phenyl)-amide (96) in pyridine (15 mL) wasadded 2.5 equivalent of freshly grounded KOH along with water (200 μL).The mixture was heated by microwave irradiation at 150° C. for 30 min,then 80-85% of the pyridine was evaporated under reduced pressure. Theresidue was poured on ice and neutralized with acetic acid. Theprecipitate was filtered off, then dried to give the title compound (1.8g, 95%): m/z=299 (M+H)⁺.

Step C: Synthesis of2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclopentanecarboxylicacid tert-butyl ester (98)

A solution of2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-hydroxycyclo-pentanecarboxylicacid tert-butyl ester (500 mg, 1.5 mmol), prepared as described inWO2005/073195,4-hydroxy-2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinoline (97, 504mg, 1.8 mmol) and triphenylphosphine (990 mg, 3.75 mmol) were stirred indry THF (40 mL) at 0° C. for 10 min. Then, DIAD (0.74 mL, 3.75 mmol) wasadded drop wise. The resulting reaction mixture was stirred at atemperature from 0° C. to 22° C. overnight. Then, volatiles wereevaporated and the residue was purified by column chromatography onsilica gel (gradient CH₂Cl₂/AcOEt, 1:0 to 95:5) to give 1.1 g (88%) ofthe title compound 98: m/z=630 (M+H)⁺.

Step D: Synthesis of2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclopentanecarboxylicacid (99)

TFA (24 mL) was added at room temperature to a solution of2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclopentanecarboxylicacid tert-butyl ester (98, 1.1 g, 1.75 mmol) and triethylsilane (510 mg,2.5 eq) in CH₂Cl₂ (24 mL). After 2 h, the reaction mixture wasconcentrated under reduced pressure, and then co-evaporated withtoluene. The residue was re-dissolved in AcOEt and successively washedwith a solution of NaHCO₃ and brine. The organic layer was dried(MgSO₄), filtered and evaporated, to give 800 mg (80%) of the titlecompound 99 (800 mg, 80%): m/z=574 (M+H)⁺.

Step E: Synthesis of1-{2-(hex-5-enylmethylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclopentanecarbonyl}amino-2-vinylcyclopropanecarboxylicacid ethyl ester (100)

A solution of2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclopentanecarboxylicacid (99, 0.77 g, 1.344 mmol), N-methylhex-5-enylamine hydrochloride(221 mg, 1.95 mmol) and diisopropylethylamine (1.17 mL, 6.72 mmol) inDMF (25 mL) was stirred at 0° C. under inert atmosphere. After 30 minHATU (741 mg, 1.95 mmol) was added and the reaction mixture was allowedto warm up to room temperature overnight. Then, DMF was evaporated andthe residue was partitioned between AcOEt and a solution of NaHCO₃.Organic layer was successively washed with water and brine, dried(MgSO₄), filtered and evaporated. The crude product was purified bysilica gel chromatography (gradient Heptane/AcOEt 80:20 to 50:50) togive 735 mg (82%) of the title compound: m/z=669 (M+H)⁺.

Step F: Synthesis of17-[2-(6-methylpyridin-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid ethyl ester (101)

1-{2-(Hex-5-enylmethylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methyl-quinolin-4-yloxy]cyclopentanecarbonyl}amino-2-vinylcyclopropanecarboxylicacid ethyl ester (100, 250 mg, 0.37 mmol) was dissolved in dry1,2-dichloroethane (250 mL). Then, nitrogen gas was bubbled through thesolution for 30 min before Hoveyda-Grubbs 2^(nd) generation (25 mg) wasadded. The resulting solution was refluxed overnight, then cooled downto room temperature and evaporated. The residue was purified by columnchromatography on silica gel (gradient AcOEt/Heptane, 3:7 to 5:5) togive 139 mg (58%) of the title compound 101.

Step G: Synthesis of17-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yl-oxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (102)

LiOH (0.42 mL, 1M) was added to a solution of17-[2-(6-methylpyridin-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo-[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid ethyl ester (101, 27 mg, 0.042 mmol) in a mixture of THF:MeOH:H₂O,2:1:1 (6 mL). The resulting solution was stirred at room temperatureovernight, then the pH was adjusted to 6 with acetic acid. The reactionmixture was successively diluted with water, extracted with CH₂Cl₂,dried (MgSO₄), filtered and evaporated to give 17 mg (65%) of the titlecompound: m/z=613 (M+H)⁺.

Step H: Synthesis of cyclopropanesulfonic acid{17-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo-[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl}-amide(103)

A mixture of the acid17-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yl-oxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (102, 28 mg, 0.046 mmol) and CDI (15 mg, 0.092 mmol) in dry THF (3mL) was heated at reflux for 2 h under nitrogen. The activation wasmonitored by LC-MS. The reaction mixture was cooled at room temperatureand cyclopropylsulfonamide (17 mg, 0.137 mmol) was added. Then, DBU (16μL, 0.105 mmol) was added and the reaction was heated at 55° C. After 24h, the pH of the reaction mixture was adjusted to 3 with citric acid(5%). Then, the solvent was evaporated, and the residue partitionedbetween AcOEt and water. The crude material was purified by preparativeHPLC to give 17 mg (52%) of the target compound 103: m/z=716 (M+H)⁺.

Example 23 Cyclopropanesulfonic acid{17-[2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carbonyl}-amide(114)

Step A: Synthesis of 2-isopropylpyridine-N-oxide (104)

A mixture of isopropylpyridine (2.1 g, 17.75 mmol) and m-CPBA (5.0 g,1.3 eq.) in CH₂Cl₂ was stirred overnight at room temperature. Then, thereaction mixture was diluted with CH₂Cl₂ (twice the volume) andsuccessively washed with aqueous sodium bicarbonate (twice) and brine,dried (Na₂SO₄) and evaporated to give 2.0 g (85%) of the title compound104.

Step B: Synthesis of 2-cyano-6-isopropylpyridine (105)

A mixture of 2-isopropylpyridine-N-oxide (104, 1.33 g, 9.7 mmol),cyanotrimethyl-silane (TMS-CN) (1.42 mL, 1.06 g, 11.0 mmol) in1,2-dichloroethane (40 mL) was stirred at room temperature for 5 min.Then, diethylcarbamoylchloride (Et₂NCOCl, 1.23 mL, 9.7 mmol) was addedand the mixture was stirred at room temperature under inert atmosphere.After 2 days, a aqueous solution of potassium carbonate (10%) was addedand the stirring was continued for 10 min. The organic layer wasseparated, and the water layer was extracted twice with1,2-dichloroethane. The combined organic layers were washed with brine,dried (Na₂SO₄) and evaporated. The residue was purified by columnchromatography on silica gel (Hexanes/AcOEt, 3:1) to give 1.06 g (74%)of the title compound: m/z=147 (M+H)⁺.

Step C: Synthesis of 6-isopropylpyridine-2-carboxylic acid (106)

A solution of 2-cyano-6-isopropylpyridine (105, 1.06 g, 7.3 mmol) in 37%aqueous HCl-MeOH (1:2) was heated to reflux overnight. Then, the solventwas evaporated, and the residue was poured into a saturated solution ofKOH. The resulting solution was refluxed overnight. Then, the solutionwas successively cooled down to room temperature and the pH of wasadjusted to 5 by addition of aqueous HCl. The resulting reaction mixturewas successively extracted with chloroform, washed with brine, dried(Na₂SO₄) and evaporated to give 0.97 g (81%) of the title compound 106:m/z=166 (M+H)⁺.

Step D: Synthesis of 6-isopropylpyridine-2-carboxylic acid(6-acetyl-3-methoxy-2-methylphenyl)amide (107)

POCl₃ (0.88 mL, 9.53 mmol) was added at −25° C. drop wise over 5 minunder nitrogen, to a stirred solution of6-isopropylpyridine-2-carboxylic acid (106, 1.43 g, 8.66 mmol) and6-acetyl-3-metoxy-2-methylaniline (1.55 g, 8.66 mmol) in dry pyridine(70 mL). The resulting solution was stirred at −10° C. for 2.5 h. Then,the reaction mixture was poured on ice, neutralized with aqueous sodiumbicarbonate and extracted 3 times with AcOEt. The organic layers werecombined, washed with brine, dried (Na₂SO₄) and evaporated. The residuewas purified by column chromatography (hexanes/AcOEt, 3:1) to give 3.54g (72%) of the title compound 107: m/z=327 (M+H)⁺.

Step E: Synthesis of4-hydroxy-2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methyl-quinoline (108)

To a solution of 6-isopropylpyridine-2-carboxylic acid(6-acetyl-3-methoxy-2-methyl-phenyl)amide (107, 0.70 g, 2.14 mmol) inpyridine (5 mL) were added 2.5 equivalents of freshly grounded KOH alongwith water (50 μL). The mixture was heated by microwave irradiation at133° C. for 55 min, then 80-85% of the pyridine was evaporated underreduced pressure. The residue was poured on ice and neutralized withacetic acid. The precipitate was filtered off, then dried to give 0.62 g(95%) of the title compound 108 (1.8 g, 95%): m/z=309 (M+H)⁺.

Step F: Synthesis of2-(1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-[2-(6-iso-propyl-pyridin-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-cyclopentanecarboxylicacid tert-butyl ester (109)

The title compound was prepared in 62% isolated yield from2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-hydroxycyclopentanecarboxylicacid tert-butyl ester and4-hydroxy-2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methylquinoline (108)following the procedure reported for the preparation2-(1-ethoxycarbonyl-2-vinylcyclopropyl-carbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclo-pentanecarboxylicacid tert-butyl ester (98): m/z=658 (M+H)⁺.

Step G: Synthesis of2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-[2-(6-iso-propyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclopentanecarboxylicacid (110)

TFA (5 mL) was added at room temperature to a solution of2-(1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-[2-(6-isopropyl-pyridin-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-cyclopentanecarboxylicacid tert-butyl ester (109, 590 mg, 0.90 mmol) and triethylsilane (280mg, 2.5 eq) in CH₂Cl₂ (5 mL). After 2 h, the reaction mixture wasconcentrated under reduced pressure to afford the desired product 110,which was used in the next step without further purifications.

Step H: Synthesis of1-{2-(hex-5-enylmethylcarbamoyl)-4-[2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclopentanecarbonyl}amino-2-vinyl-cyclopropanecarboxylicacid ethyl ester (111)

The title compound 111 was prepared in 70% isolated yield from2-(1-ethoxycarbonyl-2-vinylcyclopropylcarbamoyl)-4-[2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methyl-quinolin-4-yloxy]cyclopentanecarboxylicacid (110) following the procedure reported for the preparation of1-{2-(hex-5-enylmethylcarbamoyl)-4-[2-(6-methyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclopentanecarbonyl}amino-2-vinylcyclo-propanecarboxylicacid ethyl ester (100): m/z=697 (M+H)⁺.

Step I: Synthesis of17-[2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yl-oxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid ethyl ester (112)

1-{2-(hex-5-enylmethylcarbamoyl)-4-[2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methyl-quinolin-4-yloxy]cyclopentanecarbonyl}amino-2-vinyl-cyclopropanecarboxylicacid ethyl ester (111, 438 mg, 0.50 mmol) was dissolved in dry1,2-dichloroethane. Then, nitrogen gas was bubbled through the solutionfor 30 min before Hoveyda-Grubbs 1^(st) generation (15 mg) was added.The resulting solution was refluxed for 3 h, then more catalyst (20 mg)was added. After 2 h at reflux, another 10 mg of the catalyst was added.After 12 h at reflux, the reaction mixture was cooled down to roomtemperature. Then, scavenger MP-TMT (Agronaut Technologies Inc.) wasadded (˜300 mg) and the mixture was stirred at room temperature for 45min. The catalyst was discarded by filtration on silica gel (gradient ofCHCl₃/MeOH, 1:0 to 98:2) to give 220 mg (66%) of the title compound 112:m/z=669 (M+H)⁺.

Step J: Synthesis of17-[2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (113)

A solution LiOH (40 mg) in water (1.5 mL) was added to a solution of17-[2-(6-iso-propyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid ethyl ester (112, 220 mg, 0.33 mmol) in a mixture of MeOH (3 mL)and THF (1 mL). The resulting solution was successively heated to 55° C.for 3 h, then stirred at room temperature for 5 h. Then, the pH of thereaction mixture was adjusted to pH 6 with acetic acid and water (3 mL)was added. The resulting solution was extracted with CHCl₃. Then, theorganic layer was dried (Na₂SO₄), filtered and evaporated to give 200 mg(95%) of the title compound 113 as a white powder: m/z=641 (M+H)⁺.

Step K: Synthesis of cyclopropanesulfonic acid{17-[2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo-[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl}amide(114)

A solution of17-[2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (113, 200 mg, 0.31 mmol), DMAP (76.5 mg, 0.62 mmol), and EDC (151mg, 0.78 mmol) in DMF (5 mL) was stirred at room temperature overnight(the activation of the acid was monitored by LC-MS). Then,cyclopropylsulfonamide (191 mg, 1.56 mmol) was added, followed by DBU(228 μL, 1.56 mmol). The resulting solution was stirred overnight atroom temperature, then neutralized with acetic acid and evaporated. Theresidue was re-dissolved in MeOH and purified by preparative HPLC togive 90 mg (39%) of the title compound 114: m/z=744 (M+H)⁺.

Example 24 (6S)-Cyclopropanesulfonic acid{17-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo-[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl}amide(123) and (6R)-Cyclopropanesulfonic acid{17-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl}amide(124) Step A: Synthesis of cyclohexanecarbothioic acid amide (115)

To a suspension of cyclohexanecarboxamide (10 g, 78.6 mmol) in diethylether (300 mL) was added phosphorous pentasulfide (9.0 g, 200 mmol) inthree portions over 5 h. After stirring overnight the reaction mixturewas filtered. The mother-liquor was evaporated to give 5.5 g (49%) ofthe title compound 115.

Step B: Synthesis of 2-cyclohexylthiazole-4-carboxylic acid ethyl ester(116)

A solution of cyclohexanecarbothioic acid amide (115, 5.5 g, 38.3 mmol)and ethyl 3-bromopyruvate (90%, 8.3 g, 38.3 mmol) in THF (200 mL) washeated to reflux. After 2 h, the reaction mixture was cooled to roomtemperature for 12 h. Then, the solvent was evaporated and the residuewas purified by column chromatography (gradient of heptane/AcOEt, 90:10to 75:25) to afford 6.8 g (74%) of the title compound 116 as a clearliquid.

Step C: Synthesis of 2-cyclohexylthiazole-4-carboxylic acid (117)

To a solution of 2-cyclohexylthiazole-4-carboxylic acid ethyl ester(116, 6.8 g, 28.5 mmol) in water was added 1M LiOH (50 mL). The solutionwas kept at room temperature and monitored by LC-MS. When the hydrolysiswas completed the reaction mixture was neutralized with myriatic acidand extracted with ethyl acetate and diethyl ether. The organic phasewas dried (Na₂SO₄), filtered and concentrated under reduced pressure togive 5.0 g (83%) of the title compound 117: m/z=212 (M+H)⁺.

Step D: Synthesis of 2-cyclohexylthiazole-4-carboxylic acid(6-acetyl-3-methoxy-2-methylphenyl)amide (118)

POCl₃ (1.4 mL, 14.9 mmol) was added drop wise at −35° C. over 5 min, toa stirred solution of 2-cyclohexylthiazole-4-carboxylic acid (117, 1.5g, 7.1 mmol) and 2-acetyl-5-methoxy-6-methylaniline (1.27 g, 7.1 mmol)in dry pyridine (40 mL). After 1 h, the reaction mixture wassuccessively warmed up to room temperature for 2.5 h, evaporated andneutralized with an aqueous solution of sodium bicarbonate. Theprecipitate was filtered, washed with water and dried to give 2.6 g(95%) of the title compound 118: m/z=373 (M+H)⁺.

Step E: Synthesis of2-(2-cyclohexylthiazol-4-yl)-4-hydroxy-7-methoxy-8-methylquinoline (119)

Freshly grounded KOH (2 mmol, 112 mg) was added to a solution of2-cyclohexyl-thiazole-4-carboxylic acid(6-acetyl-3-methoxy-2-methylphenyl)amide (118, 373 mg, 2 mmol) inpyridine (20 mL). The mixture was divided into several batches and eachbatch was individually heated by microwave irradiation at 150° C. for 30min. Then, the different batches were combined and pyridine wasevaporated. The residue was treated with aqueous citric acid to give asuspension, which was subsequently diluted with a small volume of EtOH,then partitioned between water and CH₂Cl₂. Organic layer was dried(Na₂SO₄), and evaporated. The residue was purified by columnchromatography (gradient of CH₂Cl₂:MeOH, 1:0 to 93:7) to give 1.8 g(72.5%) of the title compound 119 as a white powder: m/z=355 (M+H)⁺.

Step F: Synthesis of1-{[4-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-2-(hex-5-enylmethylcarbamoyl)cyclopentanecarbonyl]amino}-2-vinylcyclo-propanecarboxylicacid ethyl ester (120)

The title compound 120 was prepared in 42% yield from1-{[4-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-2-(hex-5-enylmethylcarbamoyl)-cyclo-pentanecarbonyl]amino}-2-vinylcyclopropanecarboxylicacid ethyl ester (120) following the procedure reported for thepreparation of1-{2-(hex-5-enylmethyl-carbamoyl)-4-[2-(6-isopropyl-2-pyridyl)-7-methoxy-8-methylquinolin-4-yloxy]cyclo-pentanecarbonyl}amino-2-vinyl-cyclopropanecarboxylicacid ethyl ester (111): m/z=743 (M+H)⁺.

Step G: Synthesis of17-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid ethyl ester (121)

The title compound 121 was prepared in 50% yield from2-(2-cyclohexylthiazol-4-yl)-4-hydroxy-7-methoxy-8-methylquinoline (119)and2-(1-ethoxycarbonyl-2-vinylcyclo-propylcarbamoyl)-4-hydroxycyclopentanecarboxylicacid tert-butyl ester following the procedure reported for thepreparation of17-[2-(6-methylpyridin-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carboxylicacid ethyl ester (101): m/z=715 (M+H)⁺.

Step H: Synthesis of17-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (122)

An aqueous solution of LiOH (1M, 5 mL) was added to a solution of17-[2-(2-cyclo-hexylthiazol-4-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid ethyl ester (121) in MeOH (10 mL), THF (20 mL) and water (5 mL).The resulting solution was stirred at 50° C. for 19 h. Then, the pH ofthe reaction mixture was adjusted to 6 with myriatic acid (3M, 1.7 mL).The resulting solution was evaporated on silica and purified by columnchromatography (AcOEt/MeOH/AcOH, 74:25:1) to give 273 mg (95%) of thetitle compound 122 as a white powder: m/z=687 (M+H)⁺.

Step I: Synthesis of (6S)-cyclopropanesulfonic acid{17-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo-[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl}amide(123) and (6R)-cyclopropanesulfonic acid{17-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diaza-tricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl}amide(124)

A solution of17-[2-(2-cyclohexylthiazol-4-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (122, 173 mg, 0.25 mmol) and CDI (81 mg, 0.5 mmol) in THF (7.5 mL)was heated to reflux for 2 h (the activation of the acid was monitoredby LC-MS). Then, the reaction mixture was cooled down to roomtemperature, and cyclopropylsulfonamide (91 mg, 0.75 mmol) and DBU (8mL, 0.575 mmol) were successively added. After 12 h, the reactionmixture was neutralized with acetic acid, evaporated. The residue wasre-dissolved in water and acetonitrile, then purified by preparativeHPLC to give 21 mg (11%) of the title compound (123, first isomer):m/z=790 (M+H)⁺ and 35 mg (18%) of the second isomer 124: m/z=790 (M+H)⁺.

Example 25 Preparation ofN-[17-[2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl][1-(methyl)cyclopropyl]sulfonamide(125)

The title compound was prepared from17-[2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (65) and 1-methylcyclopropylsulfonamide following the procedurereported for the preparation ofN-[17-[8-chloro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(56): m/z=747 (M+H)⁺. ¹H NMR (CDCl₃): 0.79-0.92 (m, 2H), 1.20-2.03 (m,19H), 2.20-2.32 (m, 1H), 2.35-2.48 (m, 2H), 2.52-2.64 (m, 5H), 2.85-2.93(m, 1H), 3.04 (s, 3H), 3.05-3.14 (m, 1H), 3.35-3.46 (m, 2H), 3.97 (s,3H), 4.60 (td, J=13.2 Hz, J=2.2 Hz, 1H), 5.04 (t, J=10.5 Hz, 1H),5.30-5.47 (m, 1H), 5.61-5.69 (m, 1H), 6.30 (s, 1H), 6.32 (d, J=2.4 Hz,1H), 7.12 (d, J=9.2 Hz, 1H), 7.30 (s, 1H), 7.95 (d, J=9.0 Hz, 1H), 8.61(d, J=2.5 Hz, 1H), 10.9 (br s, 1H).

Example 26 Preparation of17-[2-(3-tert-butylpyrazol-1-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (127) Step 1: Synthesis of4-hydroxy-2-(3-tert-butylpyrazol-1-yl)-7-methoxy-8-methylquinoline (126)

The title compound was prepared from4-benzyloxy-2-chloro-7-methoxy-8-methyl-quinoline (63) and3-tert-butylpyrazole following the procedure reported for thepreparation of4-hydroxy-2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methylquinoline (64):m/z=312 (M+H)⁺.

Step 2: Synthesis of17-[2-(3-tert-butylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yl-oxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (127)

The title compound was prepared from4-hydroxy-2-(3-tert-butylpyrazol-1-yl)-7-methoxy-8-methylquinoline (126)and intermediate 26 following the procedure (Step D-F) reported for thepreparation of17-[7-methoxy-8-methyl-2-(thiazol-2-yl)quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (29): m/z=644 (M+H)⁺.

Example 27 Preparation ofN-[17-[2-(3-tert-butylpyrazol-1-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(128)

The title compound was prepared from17-[2-(3-tert-butylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (127) and cyclopropylsulfonamide following the procedure reportedfor the preparation ofN-[17-[8-chloro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(56): m/z=747 (M+H)⁺. ¹H NMR (CDCl₃): 0.95-1.12 (m, 2H), 1.13-1.30 (m,2H), 1.31-1.55 (m, 11H), 1.63-2.05 (m, 4H), 2.20-2.55 (m, 9H), 2.80-2.98(m, 1H), 3.03 (s, 3H), 3.36-3.47 (m, 2H), 3.61-3.70 (m, 1H), 3.97 (s,3H), 4.60 (t, J=12.2 Hz, 1H), 5.04 (t, J=10.3 Hz, 1H), 5.26-5.46 (m,1H), 5.61-5.69 (m, 1H), 6.35 (d, J=2.5 Hz, 1H), 6.42 (br s, 1H), 7.13(d, J=9.1 Hz, 1H), 7.32 (s, 1H), 7.95 (d, J=9.1 Hz, 1H), 8.67 (d, J=2.5Hz, 1H), 10.9 (br s, 1H).

Example 28 Preparation of17-[2-(3,5-dimethylpyrazol-1-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (130) Step 1: Synthesis of4-hydroxy-2-(3,5-dimethylpyrazol-1-yl)-7-methoxy-8-methylquinoline (129)

The title compound was prepared from4-benzyloxy-2-chloro-7-methoxy-8-methyl-quinoline (63) and3,5-dimethylpyrazole following the procedure reported for thepreparation of4-hydroxy-2-(3-isopropylpyrazol-1-yl)-7-methoxy-8-methylquinoline (64):m/z=284 (M+H)⁺.

Step 2: Synthesis of17-[2-(3,5-dimethylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (130)

The title compound was prepared from4-hydroxy-2-(3,5-dimethylpyrazol-1-yl)-7-methoxy-8-methylquinoline (129)and intermediate 26 following the procedure (Step D-F) reported for thepreparation of17-[7-methoxy-8-methyl-2-(thiazol-2-yl)quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (29): m/z=616 (M+H)⁺.

Example 29 Preparation ofN-[17-[2-(3,5-dimethylpyrazol-1-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(131)

The title compound was prepared from17-[2-(3,5-dimethylpyrazol-1-yl)-7-methoxy-8-methylquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]octadec-7-ene-4-carboxylicacid (130) and cyclopropylsulfonamide following the procedure reportedfor the preparation ofN-[17-[8-chloro-2-(4-isopropylthiazole-2-yl)-7-methoxyquinolin-4-yloxy]-13-methyl-2,14-dioxo-3,13-diazatricyclo[13.3.0.0^(4,6)]-octadec-7-ene-4-carbonyl](cyclopropyl)sulfonamide(56): m/z=719 (M+H)⁺. ¹H NMR (CDCl₃): 0.70-0.96 (m, 1H), 1.1-1.2 (m,5H), 1.4-1.55 (m, 2H), 1.80-1.93 (m, 4H), 2.15-2.25 (m, 1H), 2.30-2.40(m, 2H), 3.30 (s, 3H), 2.45-2.55 (m, 2H), 2.52 (s, 3H), 2.80 (s, 3H),2.82-2.91 (m, 2H), 3.00 (s, 3H), 3.45-3.55 (m, 2H), 3.95 (s, 3H),4.51-4.60 (m, 1H), 4.99-5.1 (m, 1H), 5.21-5.33 (m, 1H), 5.51 (m, 1H),6.00 (s, 1H), 7.03 (s, 1H), 7.10 (d, J=9.1 Hz, 1H), 7.20 (s, 1H), 7.98(d, J=9.1 Hz, 1H), 10.80 (br s, 1H).

Example 30

2-(1-Ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-pyrrolidine-1-carboxylicacid tert-butyl ester (132)

Boc-protected proline (4 g, 17.3 mmol), HATU (6.9 g, 18.2 mmol) and1-amino-2-vinyl-cyclopropanecarboxylic acid ethyl ester prepared asdescribed in WO03/099274, (3.5 g, 18.3 mmol) were dissolved in DMF (60ml) and cooled to 0° on an ice-bath. Diisopropylethyl amine (DIPEA) (6ml) was added. The ice-bath was removed and the mixture was left atambient temperature over-night. Dichloromethane (˜80 ml) was then addedand the organic phase was washed with aqueous sodium hydrogen carbonate,citric acid, water, brine and dried over sodium sulfate. Purification byflash chromatography (ether→7% methanol in ether) gave pure titlecompound (6.13 g, 96%)

Example 31

2-(1-Ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-(4-nitro-benzoyloxy)-pyrrolidine-1-carboxylicacid tert-butyl ester (133)

Compound 132 (6.13 g, 16.6 mmol), 4-nitrobenzoic acid (4.17 g, 25 mmol)and PPh₃ (6.55 g, 25 mmol) was dissolved in THF (130 ml). The solutionwas cooled to ˜0° and diisopropyl azidocarboxylate (5.1 g, 25 mmol) wasadded slowly. The cooling was then removed and the mixture was leftover-night at ambient condition. Aqueous sodium hydrogen carbonate (60ml) was added and the mixture was extracted with dichloromethane.Purification by flash chromatography (pentane-ether, 2:1→pentane-ether,1:2→2% methanol in ether) gave pure title compound (6.2 g, 72%).

Example 32

4-Nitro-benzoic acid5-(1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-pyrrolidin-3-yl ester(134)

Compound 133 (6.2 g, 12 mmol) was dissolved in an ice-cold mixture oftrifluoro-methanesulfonic acid 33% in dichloromethane. The ice-bath wasthen removed and the mixture was left at room temperature for ˜1.5 h.The solvent was evaporated and 0.25 M sodium carbonate added and themixture was extracted with dichloromethane. Evaporation gave the titlecompound (4.8 g, 95%) as a yellowish powder.

Example 33

4-Nitro-benzoic acid5-(1-ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-1-[hept-6-enyl-(4-methoxy-benzyl)-carbamoyl]-pyrrolidin-3-ylester (135)

To a solution of compound 134 (4.5 g, 10.8 mmol) in THF (160 mL) wereadded NaHCO₃ (1 tablespoon) and phosgene in toluene (1.93 M, 11.5 mL, 22mmol). The mixture was vigorously stirred for 1 h at room temperature,and then filtered and evaporated. The residue was dissolved in CH₂Cl₂(160 mL), and NaHCO₃ (1 tablespoon) andhept-5-enyl-(p-methoxybenzyl)-amine (4.3 g, 18.5 mmol) were added. Afterstirring overnight at room temperature the reaction mixture was filteredand evaporated to dryness. Flash column chromatography on silica gel(EtOAc:toluene 25:75→40:60) gave the title compound (6.59 g, 90%) as alight brown syrup.

Example 34

18-Hydroxy-14-(4-methoxy-benzyl)-2,15-dioxo-3,14,16-triaza-tricyclo[14.3.0.0*4,6]nonadec-7-ene-4-caroxylicacid ethyl ester (136)

Compound 135 (1 g, 1.48 mmol) was dissolved in 1,2-dichloroethane (2 l).The mixture was degassed for 15 min using a stream of argon.Hoveyda-Grubbs catalyst (II) (50 mg, 5 mol %) was added and the mixturewas refluxed for 4 h. The solvent was evaporated and the crude ester wasdissolved in tetrahydrofuran (100 ml), methanol (50 ml) and water (50ml). The mixture was cooled 0° C. on ice-bath. Aqueous lithium hydroxide(20 ml, 1M) was added and the mixture was stirred at 0° C. for 4 h. Thevolume was then doubled with water and the mixture acidified with aceticacid. Extraction (dichloromethane) followed by flash chromatography(methanol 1→5% in ether) gave pure title compound (450 mg, 61%).

MS (M+H)⁺ 500

Example 35

18-[2-(4-Isopropyl-thiazol-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-14-(4-methoxy-benzyl)-2,15-dioxo-3,14,16-triaza-tricyclo[14.3.0.0*4,6*]nonadec-7-ene-4-carboxylicacid ethyl ester (137)

Alcohol 136 (230 mg, 0.460 mmol), quinolinol 36 (218 mg, 0.690 mmol),and triphenylphosphine (182 mg, 0.690 mmol) were dissolved in dry THFand the mixture was cooled to 0° C. DIAD (130 μL, 0.690 mmol) was addeddropwise to the stirred solution at 0° C. during 30 minutes after whichthe solution was allowed to attain room temperature and was subsequentlystirred overnight. The solvent was evaporated and the crude material waspurified by flash column chromatography (toluene/ethyl acetate 1:1) togive the title compound (366 mg) (M+H)⁺ calcd: 796.4. found: 796.7.

Example 36

18-[2-(4-Isopropyl-thiazol-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-14-(4-methoxybenzyl)-2,15-dioxo-3,14,16-triaza-tricyclo[14.3.0.0*4,6*]nonadec-7-ene-4-carboxylicacid (138)

Ethyl ester 137 (366 mg, 0.460 mmol) was dissolved in THF/MeOH/H₂O 2:1:1(30 mL) and 1M LiOH (4.6 mL, 4.40 mmol) was added dropwise at roomtemperature during 5 minutes after which the solution was stirredovernight. The mixture was acidified to pH 3-4 by addition of solidcitric acid and the organic solvents were evaporated. The water phasewas diluted with brine (50 mL) and was then extracted trice with DCM.The combined organic phase was washed twice with brine and wasthereafter dried, filtered and concentrated. The crude was then purifiedby flash column chromatography (ethyl actetate/methanol 7:1) to give thetitle compound (212 mg, 60%). (M+H)⁺ calcd: 768.3. found: 768.7.

Example 37

1-Methyl-cyclopropanesulfonic acid[18-[2-(4-isopropyl-thiazol-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-14-(4-methoxy-benzyl)-2,15-dioxo-3,14,16-triaza-tricyclo-[14.3.0.0*4,6*]nonadec-7-ene-4-carbonyl]-amide(139)

To acid 138 (212 mg, 0.276 mmol) dissolved in dichloromethane (7 mL) wasadded EDC (69 mg, 0.359 mmol) and the reaction mixture was stirred atroom temperature. After 7 hours TLC and LC-MS indicated completeconversion of the starting material into the correspondingoxazolidinone. The reaction mixture was diluted with dichloro-methane(20 mL) and the organic phase was washed twice with water after whichthe organic phase was dried, filtered, and concentrated. The residue wasdissolved in dichloromethane (5 mL) and cyclopropylmethyl sulfonamide(53 mg, 0.394 mmol) and DBU (78 μL, 0.525 mmol) were added and thereaction mixture was stirred at room temperature for 20 hours. Themixture was diluted with dichloromethane (30 mL) and the organic phasewas washed twice with 10% citric acid and once with brine. The organicphase was dried, filtered and concentrated and the crude product waspurified by flash column chromatography (toluene/ethyl acetate 1:1, 1:2,ethyl acetate, ethyl acetate/methanol 9:1) to give the title compound(108 mg, 44%) as a colorless solid. LC-MS purity: >95%. (M+H)⁺ calcd:885.4. found: 885.7.

Example 38

1-Methyl-cyclopropanesulfonic acid{18-[2-(4-isopropyl-thiazol-2-yl)-7-methoxy-8-methyl-quinolin-4-yloxy]-2,15-dioxo-3,14,16-triaza-tricyclo[14.3.0.0*4,6*]nonadec-7-ene-4-carbonyl}-amide(140)

To compound 139 (106 mg, 0.120 mmol) dissolved in dichloromethane (18mL) were added triethylsilane (38 μL, 0.240 mmol) and TFA (9 mL) and thereaction mixture was stirred at room temperature for 1 hour. Thesolvents were evaporated and co-evaporated trice with toluene. Theresidue was dissolved in dichloromethane and the organic phase waswashed twice with saturated NaHCO₃ solution. The organic phase wasdried, filtered, and concentrated and the crude product was purified byflash column chromatography (toluene/ethyl acetate 1:1) to yield thetitle compound (73 mg, 80%) as a slightly yellow solid. LC-MSpurity: >95%. (M+H)⁺ cald: 765.3. found: 765.7.

Example 39 Alternative Route for the Preparation of Compound 34 Step A:Synthesis of 4-Amino-5-cyano-2-hydroxy-3-methylbenzoic acid ethyl ester(141)

To a solution of sodium ethoxide (1.3 L) (freshly prepared by additionof sodium metal (7.9 g, 0.35 mol) to ethanol (1.3 L)) at 0° C. was addedethylpropionyl acetate (25 g, 0.17 mol) and the solution was stirred atRT for 1 h. To the above solution was added ethoxymethylenemalononnitrile (21 g, 0.17 mol) at RT and the reaction mixture wasrefluxed at 80° C. for 2 h. The reaction mixture was cooled, neutralizedto pH=7 by addition of 1.5 N HCl and concentrated under vacuum. Theobtained residue was diluted with water (100 mL) and filtered. The solidwas washed with water and dried under vacuum at 50° C. to give the crudeproduct (27 g). The crude solid was washed with 5% ethyl acetate in pet.ether which gave pure title compound (22.5 g, 59%). TLC: EtOAc/Pet.ether, 3:7, R_(f)=0.4

Step B: Synthesis of 4-Amino-5-cyano-2-hydroxy-3-methylbenzoic acid(142)

To a solution of LiOHxH₂O (8.4 g, 0.2 mol) in ethanol/water (1:1, 300mL) was added compound 74 (22 g, 0.1 mol) at RT and the reaction mixturewas refluxed at 80° C. for 4 h. The reaction mixture was concentratedunder vacuum, the obtained residue was diluted with water (100 mL),washed with pet. ether/ethyl acetate (1:1, 2×200 mL). The aqueous layerwas separated, acidified to pH=5 using 1.5N HCl and the obtained solidproduct was filtered off. The aqueous layer was further extracted withethyl acetate (2×300 mL), dried and concentrated to give more product.The combined products was washed with 5% ethyl acetate in pet. ether togive the pure title compound (19 g, >95%).

TLC: MeOH/Chloroform, 1:4, R_(f)=0.2

Step C: Synthesis of 2-Amino-4-hydroxy-3-methylbenzonitrile (143)

A mixture of compound 75 (19 g, 0.1 mol) in quinoline (50 mL) was heatedto 170° C. for 2 h (until effervescence ceased). The reaction mixturewas cooled to RT and aqueous NaOH solution was added (1M, 500 mL)followed by pet. ether (500 mL). The reaction mixture was stirred for 15min and the aqueous layer was separated. The aqueous layer was furtherwashed with pet. ether (2×300 mL) to remove quinoline completely. Theaqueous layer was acidified with 1.5N HCl to pH=5, the solid wasfiltered off and dried under vacuum. The obtained solid was furtherwashed with 5% ethyl acetate in pet. ether to give pure title compound(12 g, 82%).

TLC: EtOAc/Pet ether, 3:7, R_(f)=0.35

Step D: Synthesis of 2-Amino-4-methoxy-3-methylbenzonitrile (144)

A mixture of compound 76 (12 g, 0.08 mol), K₂CO₃ (11 g, 0.08 mol) in dryDMF (200 mL) was stirred for 15 min at RT. To this was added MeI (13.6g, 0.096 mol) and the mixture was stirred for 4 h at RT. The reactionmixture was diluted with water (800 mL), extracted with 30% ethylacetate in pet. ether (3×300 mL). The combined organic layers werewashed with water and brine, dried and concentrated to give a crudeproduct. The crude product was washed with pet. ether to give pure titlecompound (12 g, 93%).

TLC: Pet. ether/EtOAc, 7:3, R_(f)=0.4

Step E: Synthesis of 1-(2-Amino-4-methoxy-3-methyl-phenyl)-ethanone (34)

To a solution of compound 77 (12 g, 0.074 mol) in THF (150 mL) was addedMeMgBr in diethyl ether (3M, 100 mL, 0.296 mol) at 0° C. drop-wise. Thereaction mixture was stirred at RT for 1 h and then at 55° C. for 3 h.The reaction mixture was cooled to 0° C., quenched with ice-cold 1.5NHCl till the effervescence ceases (pH=6). The reaction mixture wasdiluted with water (100 mL), extracted with ethyl acetate (2×300 mL).The combined organic layers were washed with brine, dried andconcentrated to give brown solid. The crude solid was dissolved in ethylacetate (150 mL), added pet. ether (150 mL) and passed through a bed ofsilica gel to remove color impurities and concentrated. The solidobtained was washed with 5% ethyl acetate in pet. ether which gave puretitle compound (9 g, 68%) as a yellow solid.

TLC: Pet. ether/EtOAc, 7:3, R_(f)=0.4.

Example 40 Synthesis of 3-Oxo-2-oxa-bicyclo[2.2.1]heptane-5-carboxylicacid tert-butyl ester (146)

DMAP (14 mg, 0.115 mmol) and Boc₂O (252 mg, 1.44 mmol) was added to astirred solution of 145 (180 mg, 1.15 mmol) in 2 mL CH₂Cl₂ under inertargon atmosphere at 0° C. The reaction was allowed to warm to roomtemperature and was stirred overnight. The reaction mixture wasconcentrated and the crude product was purified by flash columnchromatography (toluene/ethyl acetate gradient 15:1, 9:1, 6:1, 4:1, 2:1)which gave the title compound (124 mg, 51%) as white crystals.

¹H-NMR (300 MHz, CD₃OD) δ 1.45 (s, 9H), 1.90 (d, J=11.0 Hz, 1H),2.10-2.19 (m, 3H), 2.76-2.83 (m, 1H), 3.10 (s, 1H), 4.99 (s, 1H);¹³C-NMR (75.5 MHz, CD₃OD) δ 27.1, 33.0, 37.7, 40.8, 46.1, 81.1, 81.6,172.0, 177.7.

Alternative Method for the Preparation of Compound 146

Compound 145 (13.9 g, 89 mmol) was dissolved in dichloromethane (200 ml)and then cooled to approximately −10° C. under nitrogen. Isobutylene wasthen bubbled into the solution until the total volume had increased toapproximately 250 ml which gave a turbid solution. BF₃.Et₂O (5.6 ml,44.5 mmol, 0.5 eq.) was added and the reaction mixture was kept atapproximately −10° C. under nitrogen. After 10 min, a clear solution wasobtained. The reaction was monitored by TLC (EtOAc-Toluene 3:2 acidifiedwith a few drops of acetic acid and hexane-EtOAc 4:1, staining withbasic permanganate solution). At 70 min only traces of compound 145remained and aq. saturated NaHCO₃ (200 ml) was added to the reactionmixture, which was then stirred vigorously for 10 min. The organic layerwas washed with saturated NaHCO₃ (3×200 ml) and brine (1×150 ml), thendried with sodium sulfite, filtered and the residue was evaporated to anoily residue. Upon addition of hexane to the residue, the productprecipitated. Addition of more hexane and heating to reflux gave a clearsolution from which the product crystallized. The crystals werecollected by filtration and was washed with hexane (rt), then air-driedfor 72 h giving colourless needles (12.45 g, 58.7 mmol, 66%).

Synthesis of(1R,2R,4S)-2-((1R,2S)-1-Ethoxycarbonyl-2-vinyl-cyclopropylcarbamoyl)-4-hydroxy-cyclopentanecarboxylicacid tert-butyl ester (147)

Compound 146 (56 mg, 0.264 mmol) was dissolved in dioxane/water 1:1 (5mL) and the mixture was cooled to 0° C. 1 M lithium hydroxide (0.52 mL,0.520 mmol) was added and the mixture was stirred at 0° C. for 45minutes, after which the mixture was neutralized with 1M hydrochloricacid and evaporated and coevaporated with toluene. The crystallineresidue was dissolved in DMF (5 mL) and(1R,2S)-1-amino-2-vinyl-cyclopropane carboxylic acid ethyl esterhydrochloride (60 mg, 0.313 mmol) and diisopropylethylamine (DIEA) (138μL, 0.792 mmol) were added and the solution was cooled to 0° C. HATU(120 mg, 0.316 mmol) was added and the mixture was stirred for 0.5 h at0° C. and for an additional 2 h at room temperature. The mixture wasthen evaporated and extracted with EtOAc, washed with brine, dried,filtered and concentrated. Purification by flash column chromatography(toluene/EtOAc 1:1) provided the title compound (86 mg, 89%) as acolourless oil. The afforded oil was crystallised from ethylacetate-hexane.

Example 41 Activity of Compounds of Formula (I)

Replicon Assay

The compounds of formula (I) were examined for activity in theinhibition of HCV RNA replication in a cellular assay. The assaydemonstrated that the compounds of formula (I) exhibited activityagainst HCV replicons functional in a cell culture. The cellular assaywas based on a bicistronic expression construct, as described by Lohmannet al. (1999) Science vol. 285 pp. 110-113 with modifications describedby Krieger et al. (2001) Journal of Virology 75: 4614-4624, in amulti-target screening strategy. In essence, the method was as follows.

The assay utilized the stably transfected cell line Huh-7 luc/neo(hereafter referred to as Huh-Luc). This cell line harbors an RNAencoding a bicistronic expression construct comprising the wild typeNS3-NS5B regions of HCV type 1b translated from an Internal RibosomeEntry Site (IRES) from encephalomyocarditis virus (EMCV), preceded by areporter portion (FfL-luciferase), and a selectable marker portion(neo^(R), neomycine phosphotransferase). The construct is bordered by 5′and 3′ NTRs (non-translated regions) from HCV type 1b. Continued cultureof the replicon cells in the presence of G418 (neo^(R)) is dependent onthe replication of the HCV RNA. The stably transfected replicon cellsthat express HCV RNA, which replicates autonomously and to high levels,encoding inter alia luciferase, are used for screening the antiviralcompounds.

The replicon cells were plated in 384 well plates in the presence of thetest and control compounds which were added in various concentrations.Following an incubation of three days, HCV replication was measured byassaying luciferase activity (using standard luciferase assay substratesand reagents and a Perkin Elmer ViewLux™ ultraHTS microplate imager).Replicon cells in the control cultures have high luciferase expressionin the absence of any inhibitor. The inhibitory activity of the compoundon luciferase activity was monitored on the Huh-Luc cells, enabling adose-response curve for each test compound. EC50 values were thencalculated, which value represents the amount of the compound requiredto decrease by 50% the level of detected luciferase activity, or morespecifically, the ability of the genetically linked HCV replicon RNA toreplicate.

Inhibition Assay

The aim of this in vitro assay was to measure the inhibition of HCVNS3/4A protease complexes by the compounds of the present invention.This assay provides an indication of how effective compounds of thepresent invention would be in inhibiting HCV NS3/4A proteolyticactivity.

The inhibition of full-length hepatitis C NS3 protease enzyme wasmeasured essentially as described in Poliakov, 2002 Prot Expression &Purification 25 363 371. Briefly, the hydrolysis of a depsipeptidesubstrate, Ac-DED(Edans)EEAbuψ[COO]ASK(Dabcyl)-NH₂ (AnaSpec, San José,USA), was measured spectrofluorometrically in the presence of a peptidecofactor, KKGSVVIVGRIVLSGK (Åke Engström, Department of MedicalBiochemistry and Microbiology, Uppsala University, Sweden). [Landro,1997 #Biochem 36 9340-9348]. The enzyme (1 nM) was incubated in 50 mMHEPES, pH 7.5, 10 mM DTT, 40% glycerol, 0.1% n-octyl-D-glucoside, with25 μM NS4A cofactor and inhibitor at 30° C. for 10 min, whereupon thereaction was initiated by addition of 0.5 μM substrate. Inhibitors weredissolved in DMSO, sonicated for 30 sec. and vortexed. The solutionswere stored at −20° C. between measurements.

The final concentration of DMSO in the assay sample was adjusted to3.3%. The rate of hydrolysis was corrected for inner filter effectsaccording to published procedures. [Liu, 1999 Analytical Biochemistry267 331-335]. Ki values were estimated by non-linear regression analysis(GraFit, Erithacus Software, Staines, MX, UK), using a model forcompetitive inhibition and a fixed value for Km (0.15 μM). A minimum oftwo replicates was performed for all measurements.

The following Table 1 lists compounds that were prepared according toany one of the above examples. The activities of the compounds testedare also depicted in Table 1

EC₅₀ (μM) Ki (μM) Compound nr. structure Replicon assay Enzymatic assay 29

10      —  47

0.00618  0.00050  46

0.91    —  91

8.54 × 10⁻³ 5.00 × 10⁻⁵  55

 0.36743075 5.00 × 10⁻³  81

10      1      82

8.321539  9.40 × 10⁻³  56

2.93 × 10⁻³ 1.00 × 10⁻⁴  57

1.87 × 10⁻³ 3.00 × 10⁻⁴  94

3.26 × 10⁻³ 1.00 × 10⁻⁴  48

2.33 × 10⁻³ 2.50 × 10⁻⁴  95

4.04 × 10⁻³ 1.00 × 10⁻⁴  75

5.75 × 10⁻² —  71

10      — 103

6.30 × 10⁻³ —  72

6.60 × 10⁻² —  66

0.0036   —

Example 32 In Vivo Effects of Ritonavir on the Pharmacokinetics ofCompound Nr. 47 in Rat

Oral pharmacokinetics of Compound nr. 47 in male and femaleSprague-Dawley rats after a single dose at 10 mg/kg, using a formulationin 50% PEG400/water and the influence of “boosting” with 10 mg/kgritonavir were investigated.

Four male and four female Sprague-Dawley (SD)-rats (approx. body weight200-250 g) were randomly divided into 2 groups of 2 males and femaleseach (boosted and non-boosted) based on body weight. The weight of theindividual animals did not differ too much from the group mean. Theanimals were fasted shortly before the trial. Drinking water remainedavailable ad libitum.

Rats from the non-boosted group received a single oral 10 mg/kg dose ofCompound nr. 47, formulated as a 3 mg/ml 50% PEG400/water at pH 8. Ratsfrom the boosted group received a single oral dose of ritonavir, about30 minutes before single oral dosing with 10 mg/kg of Compound nr. 47.The drug formulations were administered by oral gavage.

From each rat, a 0.5 ml blood sample was collected at 0.5 h, 1 h, 2 h, 4h and 8 h after dosing. Plasma concentrations were determined usingHPLC-MS. Results are shown in the table 2 below, expressed as foldchange in pharmacokinetic parameter of the boosted group as compared tothe non-boosted group.

TABLE 2 pharmacokinetic parameter ritonavir Compound nr. 47 C_(max) 2.2AUC 2.5

These results demonstrate that ritonavir substantially enhances thepharmacokinetics of Compound nr. 47 in rat, with overall exposuresexpressed as AUC increasing over 2-fold.

The invention claimed is:
 1. A combination comprising: (a) a compoundhaving the formula

or an N-oxide, a pharmaceutically acceptable salt, or stereoisomerthereof; and (b) another anti-HCV compound.
 2. The combination accordingto claim 1 other than an N-oxide or pharmaceutically acceptable salt. 3.The combination according to claim 1 other than an N-oxide.
 4. Thecombination as claimed in claim 1, wherein the other anti-HCV compoundis an HCV polymerase inhibitor, an HCV protease inhibitor, an inhibitorof another target in the HCV life cycle, an immunomodulatory agent, anantiviral agent, or combinations thereof.
 5. The combination as claimedin claim 4, wherein the other anti-HCV compound is an HCV polymeraseinhibitor.
 6. The combination as claimed in claim 4, for simultaneous,separate, or sequential use in the treatment of HCV infections.
 7. Thecombination as claimed in claim 5, for simultaneous, separate, orsequential use in the treatment of HCV infections.
 8. A method ofinhibiting HCV replication comprising administering to a patient in needthereof, a compound of the formula

in combination with another compound.
 9. The method as claimed in claim8, wherein the other compound is another anti-HCV compound.
 10. Themethod as claimed in claim 9, wherein the other anti-HCV compound is anHCV polymerase inhibitor, an HCV protease inhibitor, an inhibitor ofanother target in the HCV life cycle, an immunomodulatory agent, anantiviral agent, or combinations thereof.
 11. The method as claimed inclaim 9, wherein the other anti-HCV compound is an HCV polymeraseinhibitor.
 12. The method as claimed in claim 10, wherein thecombination is administered simultaneously, separately, or sequentiallyin the treatment of the HCV replication inhibition.
 13. The method asclaimed in claim 11, wherein the combination is administeredsimultaneously, separately, or sequentially in the treatment of the HCVreplication inhibition.
 14. A pharmaceutical composition comprising acarrier, and as active ingredient an anti-virally effective amount of acombination comprising (a) a compound having the formula

or an N-oxide, a pharmaceutically acceptable salt, or stereoisomerthereof; and (b) another anti-HCV compound.
 15. The pharmaceuticalcomposition of claim 14 wherein the other anti-HCV compound is an HCVpolymerase inhibitor, an HCV protease inhibitor, an inhibitor of anothertarget in the HCV life cycle, an immunomodulatory agent, an antiviralagent, or combinations thereof.
 16. The pharmaceutical composition ofclaim 14 wherein the other anti-HCV compound is an HCV polymeraseinhibitor.
 17. The combination of claim 4, wherein the other anti-HCVcompound is a protease inhibitor.
 18. The method of claim 8, wherein theother compound is a protease inhibitor.
 19. The pharmaceuticalcomposition of claim 14, wherein the other anti-HCV compound is aprotease inhibitor.